From e8c6120b48683bb156dad22291a02807f069fab7 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 7 Feb 2023 09:38:49 +0100
Subject: [PATCH 001/124] Fixed ##Enriched Pathways problem in report

---
 assets/RNAseq_report.Rmd | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 779af63c..70d26614 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1883,7 +1883,7 @@ Inside the pathway analysis results folder, a subfolder for each contrast used f
 </details><br>",
 gost_text,
 
-"
+"\n
 ## Enriched pathways
 The plot below summarizes the pathways that were
 found significantly enriched in DE genes for each contrast (padj value < ", pval_text, ").

From f0829f561b3c179f391934558104bbd52d27d82d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 7 Feb 2023 09:42:06 +0100
Subject: [PATCH 002/124] Updated changelog

---
 CHANGELOG.md | 11 +++++++++++
 1 file changed, 11 insertions(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 70059f9b..16c51d05 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -3,6 +3,17 @@
 The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
 and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
 
+## 2.1dev
+
+### Added
+
+### Changed
+
+### Fixed
+
+- [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
+
+
 ## 2.1 - Wheat Fields
 
 ### Added

From db1732c5efeea187cf32dd8675d500d671144332 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 7 Feb 2023 09:56:48 +0100
Subject: [PATCH 003/124] prettier

---
 CHANGELOG.md | 1 -
 1 file changed, 1 deletion(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 16c51d05..2ea1ab61 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -13,7 +13,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 - [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
 
-
 ## 2.1 - Wheat Fields
 
 ### Added

From 83bffa388a5f7cd124114fcefc953ed303b3bfcc Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 9 Feb 2023 10:15:32 +0100
Subject: [PATCH 004/124] Added optional --quote

---
 assets/RNAseq_report.Rmd | 13 +++++++++++--
 bin/Execute_report.R     |  2 ++
 conf/test.config         |  1 +
 conf/test_full.config    |  1 +
 modules/local/report.nf  |  5 ++++-
 nextflow.config          |  1 +
 nextflow_schema.json     |  4 ++++
 7 files changed, 24 insertions(+), 3 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 779af63c..dff741d6 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -43,6 +43,7 @@ params:
 
     #Additional args for the report
     path_proj_summary: ''
+    path_quote: ''
     path_software_versions: ''
     revision: ''
 
@@ -1976,8 +1977,16 @@ and the subfolder: `pathway_heatmaps`.
 
 # Summary and outlook
 
-\
-The results for all work packages, as described in the quote can be found in this report.
+```{r quote, echo=FALSE, results='asis'}
+# Only add hyperlink to quote file if it was provided in the params
+quote_string <- ifelse(
+    isProvided(params$path_quote),
+    paste0("The results for all work packages, as described in the [quote](", params$path_quote, "), can be found in this report."),
+    paste0("The results for all work packages, as described in the quote, can be found in this report.")
+)
+cat(quote_string)
+```
+
 Further support for this project will be restricted to the results presented in this report.
 For further analysis (e.g. the re-analysis of the dataset) we will generate a new quote containing cost estimates.
 
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index a67ccfa0..91125536 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -34,6 +34,7 @@ option_list = list(
     make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"),
 
     make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"),
+    make_option(c("--path_quote"), type="character", default=NULL, help="Path to the quote PDF", metavar="character"),
     make_option(c("-v", "--software_versions"), type="character", default=NULL, help="Versions file", metavar="character"),
     make_option(c("-j", "--revision"), type="character", default=NULL, help="Rnadeseq workflow revision", metavar="character"),
 
@@ -79,6 +80,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 min_DEG_pathway = opt$min_DEG_pathway,
 
                                 path_proj_summary = opt$proj_summary,
+                                path_quote = opt$path_quote,
                                 path_software_versions = opt$software_versions,
                                 revision = opt$revision,
 
diff --git a/conf/test.config b/conf/test.config
index 72db4f7c..05a27ad8 100644
--- a/conf/test.config
+++ b/conf/test.config
@@ -29,4 +29,5 @@ params {
     multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip'
     run_pathway_analysis = true
     genome = 'GRCm38'
+    quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf'
 }
diff --git a/conf/test_full.config b/conf/test_full.config
index 8052eb5f..66cc60f9 100644
--- a/conf/test_full.config
+++ b/conf/test_full.config
@@ -28,4 +28,5 @@ params {
     multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip'
     run_pathway_analysis = true
     genome = 'GRCm38'
+    quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf'
 }
diff --git a/modules/local/report.nf b/modules/local/report.nf
index a4ead56f..bbaeff6b 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -35,7 +35,9 @@ process REPORT {
     def round_DE_opt = params.round_DE ? "--round_DE $params.round_DE" : ''
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
-    def custom_gmt_opt = custom_gmt.name  != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
+    def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
+    
+    def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
 
     def citest_opt = params.citest ? "--citest TRUE" : ''
 
@@ -70,6 +72,7 @@ process REPORT {
         --species_library $params.species_library \
         --keytype $params.keytype \
         --min_DEG_pathway $params.min_DEG_pathway \
+        $quote_opt \
         --proj_summary $proj_summary \
         --software_versions $software_versions \
         --revision $workflow.manifest.version \
diff --git a/nextflow.config b/nextflow.config
index dd6cccae..f430db4c 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -42,6 +42,7 @@ params {
     // Additional args for the report
     project_summary = null
     multiqc = 'NO_FILE4'
+    quote = 'NO_FILE5'
 
     // Additional arg to indicate that a github test is run
     citest = false
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 5d891d36..ed2ad81d 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -149,6 +149,10 @@
                 "multiqc": {
                     "type": "string",
                     "description": "multiqc.zip folder containing the multiQC plots and report."
+                },
+                "quote": {
+                    "type": "string",
+                    "description": "Quote file to link in the report‚."
                 }
             }
         },

From e8a2a2f0187d4925ed250970f14454b709c4c865 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 9 Feb 2023 10:23:41 +0100
Subject: [PATCH 005/124] changed container to dev

---
 .github/workflows/ci.yml | 8 ++++----
 Dockerfile               | 6 +++---
 environment.yml          | 2 +-
 modules/local/report.nf  | 2 +-
 4 files changed, 9 insertions(+), 9 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index e3e9eca2..f6ee4de7 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -36,13 +36,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:2.1
+        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:2.1
+          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
@@ -88,13 +88,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:2.1
+        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:2.1
+          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
diff --git a/Dockerfile b/Dockerfile
index 5d413a56..aef20cc2 100644
--- a/Dockerfile
+++ b/Dockerfile
@@ -3,14 +3,14 @@ LABEL authors="Gisela Gabernet, Alexander Peltzer" \
     description="Docker image containing all requirements for qbic-pipelines/rnadeseq pipeline"
 COPY environment.yml /
 #RUN conda install -c conda-forge mamba
-RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.1 && \
+RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \
     mamba clean --all --yes
 RUN apt-get update -qq && \
     apt-get install -y zip procps ghostscript
 # Add conda installation dir to PATH
-ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.1/bin:$PATH
+ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH
 # Dump the details of the installed packates to a file for posterity
-RUN mamba env export --name qbic-pipelines-rnadeseq-2.1 > qbic-pipelines-rnadeseq-2.1.yml
+RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml
 # Instruct R processes to use these empty files instead of clashing with a local config
 RUN touch .Rprofile
 RUN touch .Renviron
diff --git a/environment.yml b/environment.yml
index a7ce1ce0..60d64f8b 100644
--- a/environment.yml
+++ b/environment.yml
@@ -1,7 +1,7 @@
 # You can use this file to create a conda environment for this pipeline:
 #   conda env create -f environment.yml
 # use this to find packages: https://anaconda.org/
-name: qbic-pipelines-rnadeseq-2.1
+name: qbic-pipelines-rnadeseq-dev
 channels:
   - bioconda
   - conda-forge
diff --git a/modules/local/report.nf b/modules/local/report.nf
index a4ead56f..9fa0858a 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -1,6 +1,6 @@
 process REPORT {
 
-    container 'qbicpipelines/rnadeseq:2.1'
+    container 'qbicpipelines/rnadeseq:dev'
 
     input:
     path gene_counts

From da4e786a77a7f0c84522055ceed8b14f64137bd4 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 9 Feb 2023 11:22:28 +0100
Subject: [PATCH 006/124] Added gprofiler version string to report

---
 assets/RNAseq_report.Rmd | 36 ++++--------------------------------
 1 file changed, 4 insertions(+), 32 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 779af63c..95fd2a41 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1590,33 +1590,7 @@ if (isProvided(params$custom_gmt)){
             print(gmt_url)
             print("Got error:")
             print(gost_error)
-            print("For reproducibility reasons, try to download the GMT file manually by visiting https://biit.cs.ut.ee/gprofiler/gost, then selecting the correct organism and finally, in datasources, clicking 'combined ENSG.gmt'. Then provide it to the pipeline with the parameter `--custom_gmt`")
-        }
-    )
-
-    # Also try to get the version info from the current build
-    gpro_version_url <- paste0("https://biit.cs.ut.ee/gprofiler/api/util/data_versions?organism=", organism)
-    gpro_version_string <- ""
-    tryCatch(
-        {
-            gpro_command <- paste0("curl ", gpro_version_url)
-            sys_return <- system(gpro_command)
-            if ("gprofiler_version" %in% sys_return) {
-                info <- sys_return
-                sources <- info$sources
-                gpro_version_string <- paste0("`gprofiler: ", info$gprofiler_version, "`, ")
-            } else {
-                print("Failed to obtain the gprofiler version with this command:")
-                print(gpro_command)
-                print("For reproducibility reasons, find the gprofiler command on https://biit.cs.ut.ee/gprofiler/gost.")
-            }
-        },
-        error=function(version_error) {
-            print("Failed to fetch the gprofiler version with this command:")
-            print(gpro_command)
-            print("Got error:")
-            print(version_error)
-            print("For reproducibility reasons, find the gprofiler command on https://biit.cs.ut.ee/gprofiler/gost.")
+            print("For reproducibility reasons, please try to download the GMT file manually by visiting https://biit.cs.ut.ee/gprofiler/gost, then selecting the correct organism and finally, in datasources, clicking 'combined ENSG.gmt'. Then provide it to the pipeline with the parameter `--custom_gmt`")
         }
     )
 }
@@ -2042,14 +2016,12 @@ if (isProvided(params$custom_gmt)){
     database_string <- paste0(database_string, "`", dbs, "`.")
 } else {
     database_string = "Also, the following databases were queried: "
-    if (gpro_version_string != "") {
-        database_string <- paste0(database_string, gpro_version_string, ",")
-    }
     database_version_info <- get_version_info(params$organism)
+    database_string <- paste0(database_string, "`gprofiler: ", database_version_info$gprofiler_version, "`")
     for (db in datasources) {
-        database_string <- paste0(database_string, "`", db, ": ", gsub("\n", ", ", database_version_info$sources[[db]]$version), "`, ")
+        database_string <- paste0(database_string, ", `", db, ": ", database_version_info$sources[[db]]$version, "`")
     }
-    database_string <- paste0(strsplit(database_string, ",\\s*(?=[^,]+$)", perl=TRUE)[[1]], ".")
+    database_string <- paste0(database_string, ".") # Add final dot to sentence
 }
 
 cat(paste0("For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2," `, `AnnotationDbi v", version_annotation, "` and `", name_species, " v", version_annotation, "` were employed. ", database_string))

From 3bfbe1747f4529ee4b38e1452f49813a74094b11 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 9 Feb 2023 11:31:48 +0100
Subject: [PATCH 007/124] rm trailing whitespace

---
 modules/local/report.nf | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/modules/local/report.nf b/modules/local/report.nf
index bbaeff6b..d131c588 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -36,7 +36,7 @@ process REPORT {
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
-    
+
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
 
     def citest_opt = params.citest ? "--citest TRUE" : ''

From b2092a3b922cdd78ffdd896bde557ad3572bd82d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 13 Feb 2023 10:32:25 +0100
Subject: [PATCH 008/124] added some missing SVG calls

---
 .github/workflows/ci.yml |  8 ++++----
 Dockerfile               |  6 +++---
 assets/RNAseq_report.Rmd | 22 ++++++++++++++++++++--
 environment.yml          |  4 +++-
 modules/local/report.nf  |  2 +-
 5 files changed, 31 insertions(+), 11 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index e3e9eca2..f6ee4de7 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -36,13 +36,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:2.1
+        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:2.1
+          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
@@ -88,13 +88,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:2.1
+        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:2.1
+          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
diff --git a/Dockerfile b/Dockerfile
index 5d413a56..aef20cc2 100644
--- a/Dockerfile
+++ b/Dockerfile
@@ -3,14 +3,14 @@ LABEL authors="Gisela Gabernet, Alexander Peltzer" \
     description="Docker image containing all requirements for qbic-pipelines/rnadeseq pipeline"
 COPY environment.yml /
 #RUN conda install -c conda-forge mamba
-RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.1 && \
+RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \
     mamba clean --all --yes
 RUN apt-get update -qq && \
     apt-get install -y zip procps ghostscript
 # Add conda installation dir to PATH
-ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.1/bin:$PATH
+ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH
 # Dump the details of the installed packates to a file for posterity
-RUN mamba env export --name qbic-pipelines-rnadeseq-2.1 > qbic-pipelines-rnadeseq-2.1.yml
+RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml
 # Instruct R processes to use these empty files instead of clashing with a local config
 RUN touch .Rprofile
 RUN touch .Renviron
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 779af63c..7aff6940 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -218,6 +218,9 @@ invisible( lapply(c(
 "tools",
 "yaml"
 ), library, character.only=T))
+
+# Import python sys so that the save_image command works
+reticulate::py_run_string("import sys")
 ```
 
 ```{r create_outdirs, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
@@ -1037,8 +1040,9 @@ sampleDistMatrix <- as.matrix(sampleDists)
 colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255)
 par(oma=c(3,3,3,3))
 dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey", fontsize_row=label_size(nrow(sampleDistMatrix)), fontsize_col=label_size(ncol(sampleDistMatrix)), show_rownames=label_size(nrow(sampleDistMatrix))>=4, show_colnames=label_size(ncol(sampleDistMatrix))>=4)
-export(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.png")
-export(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.pdf")
+save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.pdf")
+save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.png")
+save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.svg")
 #dend <- hclust(dist(mtcars, method = "euclidean"), method = "complete")
 #capture.output(cutree(dend, k = 3))
 #write.table(make_row_col_mat(sampleDistMatrix), file="differential_gene_expression/plots/Heatmaps_of_distances_index.tsv", row.#names=F, quote=F)
@@ -1696,6 +1700,9 @@ for (file in contrast_files){
         ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.png"),
             device="png",
             height=10, width=15, units="cm", dpi=300, limitsize=F)
+        ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.svg"),
+            device="svg",
+            height=10, width=15, units="cm", dpi=300, limitsize=F)
     }
     # Remove parents column to be able to save the table in tsv format
     pathway_gostres_table <- pathway_gostres
@@ -1739,6 +1746,7 @@ for (file in contrast_files){
                 xlab("") + ylab("Gene fraction (DE genes / Pathway size)")
             ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname, "_", db_source, "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F)
             ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F)
+            ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F)
 
             # Plotting heatmaps and KEGG pathways for all pathways
             print("Plotting heatmaps...")
@@ -1765,6 +1773,11 @@ for (file in contrast_files){
                         pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""))
                         pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
                         dev.off()
+                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""))
+                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+                        dev.off()
+                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+                        dev.off()
                         write.table(make_row_col_mat(mat), file=paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, "_index.tsv", sep=""), row.names=F, quote=F)
                     }
 
@@ -1847,6 +1860,11 @@ if (isProvided(params$path_genelist)){
         pdf(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf", sep=""))
         pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
         dev.off()
+        svg(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg", sep=""))
+        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+        dev.off()
+        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+        dev.off()
         write.table(make_row_col_mat(mat), file=paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list_index.tsv", sep=""), row.names=F, quote=F)
     }
 }
diff --git a/environment.yml b/environment.yml
index a7ce1ce0..abb9e9cc 100644
--- a/environment.yml
+++ b/environment.yml
@@ -21,7 +21,8 @@ dependencies:
   - bioconda::bioconductor-tximport=1.22.0
   - bioconda::bioconductor-vsn=3.62.0
   - conda-forge::pandoc=2.17.1.1
-  - conda-forge::phantomjs=2.1.1 #this is necessary for exporting heatmaply plots
+  - conda-forge::plotly=5.13.0 #this is necessary for exporting plots with save_image()
+  - conda-forge::python-kaleido=0.2.1 #this is necessary for exporting plots with save_image()
   - conda-forge::r-base=4.1.2
   - conda-forge::r-details=0.3.0
   - conda-forge::r-dplyr=1.0.10
@@ -40,6 +41,7 @@ dependencies:
   - conda-forge::r-plyr=1.8.6
   - conda-forge::r-rcolorbrewer=1.1_2
   - conda-forge::r-reshape2=1.4.4
+  - conda-forge::r-reticulate=1.28 #this is necessary for exporting heatmaply plots with save_image()
   - conda-forge::r-rmarkdown=2.11
   - conda-forge::r-sessioninfo=1.2.2
   - conda-forge::r-svglite=2.1.0
diff --git a/modules/local/report.nf b/modules/local/report.nf
index a4ead56f..9fa0858a 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -1,6 +1,6 @@
 process REPORT {
 
-    container 'qbicpipelines/rnadeseq:2.1'
+    container 'qbicpipelines/rnadeseq:dev'
 
     input:
     path gene_counts

From 01f4bda9e5829c49e882a49c4ba6a9b7c987893a Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 13 Feb 2023 15:08:48 +0100
Subject: [PATCH 009/124] Changed text for quote/offer in report

---
 assets/RNAseq_report.Rmd | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index dff741d6..4ce2f992 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1981,8 +1981,8 @@ and the subfolder: `pathway_heatmaps`.
 # Only add hyperlink to quote file if it was provided in the params
 quote_string <- ifelse(
     isProvided(params$path_quote),
-    paste0("The results for all work packages, as described in the [quote](", params$path_quote, "), can be found in this report."),
-    paste0("The results for all work packages, as described in the quote, can be found in this report.")
+    paste0("The results for all work packages, as described in the [quote/offer](", params$path_quote, "), can be found in this report."),
+    paste0("The results for all work packages (no quote/offer given to describe them) can be found in this report.")
 )
 cat(quote_string)
 ```

From b37a605d8e0bed53c003e2840c9e33c331270ab2 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 13 Feb 2023 15:59:26 +0100
Subject: [PATCH 010/124] Updated 5sum for rsem

---
 tests/test_star_rsem.yml | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 1df704ee..dc0c6683 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -4,11 +4,11 @@
     - test_star_rsem
   files:
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
-      md5sum: 60aff9f171a58cd2c9db9e60eda3a506
+      md5sum: 216106ef902281c5d7c72715583e384c
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
       md5sum: 7a4db95674a9c04aaafaef339edcfcc7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_MCF7_vs_GM12878.tsv
-      md5sum: b7e914acf8a0d379e0ace343cfd1a69b
+      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: f1825ed59878115a279db7151b0ed0a1
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/CXCL2_ENSG00000081041.pdf

From babba2e5a2c3661a7a6cfa40470c7a58d1415c5a Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Feb 2023 14:10:50 +0100
Subject: [PATCH 011/124] removed rsem/salmon files from output.zip, instead
 added txt file which lists the file names --> also added to the pytests

---
 assets/RNAseq_report.Rmd   | 16 ++++++++--------
 tests/test_star_rsem.yml   |  2 ++
 tests/test_star_salmon.yml |  2 ++
 3 files changed, 12 insertions(+), 8 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 70d26614..e4a20bbd 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -454,8 +454,8 @@ if (params$input_type == "featurecounts") {
     tx2gene_gtf <- distinct(tx2gene_gtf)
 
     # As the tximport parameter ignoreTxVersion=T does not reliably work (I think it only ignores the version
-    #in the input files, not in the gtf), this removes decimals from the transcript_id values,
-    #e.g. transcript_id "AT1G01010.1" --> transcript_id "AT1G01010"
+    # in the input files, not in the gtf), this removes decimals from the transcript_id values,
+    # e.g. transcript_id "AT1G01010.1" --> transcript_id "AT1G01010"
     tx2gene_gtf[] <- lapply(tx2gene_gtf, function(x) gsub("\\.\\d+", "", x))
     colnames(tx2gene_gtf) <- c("transcript_id", "gene_id") #, "TXID"
     gene_names <- gtf[c("gene_id", "gene_name")]
@@ -463,7 +463,7 @@ if (params$input_type == "featurecounts") {
     gene_names <- distinct(gene_names)
     rownames(gene_names) <- gene_names[,1]
 
-## Load Rsem count table
+## Load Rsem count files
     if (params$input_type == "rsem") {
 
         #rsem saves output into one file per sample in an output folder
@@ -555,11 +555,11 @@ write.table(sizeFactors(cds),paste("differential_gene_expression/gene_counts_tab
 # Write cds assay table to file
 write.table(counts(cds, normalized=T), paste("differential_gene_expression/gene_counts_tables/deseq2_table.tsv", sep=""), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
 
-if (params$input_type %in% c("rsem", "salmon")) {
-
-    # Else copy raw count sample files/folders
-    system("mkdir differential_gene_expression/gene_counts_tables/raw_gene_counts/")
-    system(paste0("cp -r ",params$path_gene_counts,"/* differential_gene_expression/gene_counts_tables/raw_gene_counts/"))
+# Or for rsem/salmon save folder/file names to txt file
+if (params$input_type == "rsem") {
+    write(files, file="differential_gene_expression/metadata/star_rsem_files.txt")
+} else if (params$input_type == "salmon") {
+    write(files, file="differential_gene_expression/metadata/star_salmon_files.txt")
 }
 
 ############################## TRANSFORMED AND NORMALIZED COUNTS ###################
diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 1df704ee..2c75d362 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -3,6 +3,8 @@
   tags:
     - test_star_rsem
   files:
+    - path: results_test/differential_gene_expression/metadata/star_salmon_files.txt
+      md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
       md5sum: 60aff9f171a58cd2c9db9e60eda3a506
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index 03ee8df6..8b424779 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -3,6 +3,8 @@
   tags:
     - test_star_salmon
   files:
+    - path: results_test/differential_gene_expression/metadata/star_salmon_files.txt
+      md5sum: db1da70879624425d9908b6679572f70
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
       md5sum: c80564418790c6d4a60a460f594b2934
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv

From 7d3f402a077e34aac4ee4d0fd942a79256b75fbf Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Feb 2023 15:54:33 +0100
Subject: [PATCH 012/124] Updated documentation (also in report)

---
 assets/RNAseq_report.Rmd | 11 ++++++++++-
 docs/output.md           |  3 ++-
 2 files changed, 12 insertions(+), 2 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index e4a20bbd..587604ad 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1053,6 +1053,14 @@ dist_heatmap %>% layout(height=700, width=800) %>% config(modeBarButtonsToAdd =
 
 After unzipping the results ZIP file, the differential expression results can be found in the directory [differential_gene_expression/](./differential_gene_expression), containing the following subfolders:
 
+```{r aligner_strings, echo=FALSE, message=FALSE, warning=FALSE}
+
+# Change the different strings depending on which input_type was used
+    featurecounts_string <- ifelse(params$input_type == "featurecounts", "
+    - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline; used for the differential gene expression analysis.", "")
+    rsem_salmon_string <- ifelse(params$input_type %in% c("rsem", "salmon"), paste0("- `star_", params$input_type, "_files.txt`: list of ", ifelse(params$input_type == "rsem", "RSEM counts files", "Salmon quantification files"), " that were provided as input; used for the differential gene expression analysis."), "")
+```
+
 <br><details>
 <summary>Expand/collapse</summary>
 - `metadata/`
@@ -1060,8 +1068,9 @@ After unzipping the results ZIP file, the differential expression results can be
     - `contrasts.tsv`: contrasts used for DE analysis (if provided).
     - `linear_model.txt`: linear model used for DE analysis.
     - `gene_list.txt`: provided list of interesting genes (if provided).
+    `r rsem_salmon_string`
 - `gene_counts_tables/`
-    - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline and used for the differential gene expression analysis.
+    `r featurecounts_string`
     - `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts.
     - `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation.
     - `sizeFactor_libraries.tsv`: size factors for each sample.
diff --git a/docs/output.md b/docs/output.md
index cb85112a..ff953f84 100644
--- a/docs/output.md
+++ b/docs/output.md
@@ -26,8 +26,9 @@ This directory contains the zipped results. When unzipping them, the following s
   - `contrasts.tsv`: contrasts used for DE analysis (if provided).
   - `linear_model.txt`: linear model used for DE analysis.
   - `gene_list.txt`: provided list of interesting genes (if provided).
+  - `star_[rsem/salmon]_files.txt`: list of counts files that were provided as input (only if `--input_type` is `rsem` or `salmon`).
 - `gene_counts_tables/`
-  - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline and used for the differential gene expression analysis.
+  - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline and used for the differential gene expression analysis (only if `--input_type` is `featurecounts`).
   - `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts (only when rlog transformation is used).
   - `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation (only when vst transformation is used).
   - `sizeFactor_libraries.tsv`: size factors for each sample.

From c7f7c29fdbd1dfbbe28b42e34ca7ffa9691d3b33 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 16 Feb 2023 07:56:01 +0100
Subject: [PATCH 013/124] Fixed rsem test

---
 tests/test_star_rsem.yml | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 2c75d362..30e669b6 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -3,7 +3,7 @@
   tags:
     - test_star_rsem
   files:
-    - path: results_test/differential_gene_expression/metadata/star_salmon_files.txt
+    - path: results_test/differential_gene_expression/metadata/star_rsem_files.txt
       md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
       md5sum: 60aff9f171a58cd2c9db9e60eda3a506

From ea3ce9b36fe52bf1829e1c36ea685912e30a0e66 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 20 Feb 2023 15:04:49 +0100
Subject: [PATCH 014/124] Heatmap sizes scale depending on data

---
 CHANGELOG.md             | 16 +++++++++++++++
 assets/RNAseq_report.Rmd | 42 +++++++++++++---------------------------
 2 files changed, 29 insertions(+), 29 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 70059f9b..041455e9 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -3,6 +3,22 @@
 The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
 and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
 
+## 2.1.1 - Wheat Fields hotfix I
+
+### Added
+
+- [#197](https://github.com/qbic-pipelines/rnadeseq/pull/197) Added gprofiler version string to report
+- [#196](https://github.com/qbic-pipelines/rnadeseq/pull/196) Added optional `--quote` parameter
+
+### Changed
+
+- [#198](https://github.com/qbic-pipelines/rnadeseq/pull/198) Changed heatmaps to scale in size automatically
+
+### Fixed
+
+- [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed format error in report pathway section
+
+
 ## 2.1 - Wheat Fields
 
 ### Added
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 7aff6940..09752e5d 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -83,13 +83,6 @@ label_size <- function(dimension_int) {
     val <- max(c(2, 10-dimension_int/15))
     return(val)
 }
-
-# Function for making a dataframe of row/column names to save to a file in case the plot labels are not readable
-make_row_col_mat <- function(mat) {
-    row_col_max_length <- max(c(length(rownames(mat)), length(colnames(mat))))
-    return(data.frame(Row.Names=c(rownames(mat), rep(NA, row_col_max_length-length(rownames(mat)))),
-                          Column.Names=c(colnames(mat), rep(NA, row_col_max_length-length(colnames(mat))))))
-}
 ```
 
 ```{r param_validation, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
@@ -1039,13 +1032,10 @@ sampleDists <- dist(t(assay(if (params$rlog) rld else vsd)))
 sampleDistMatrix <- as.matrix(sampleDists)
 colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255)
 par(oma=c(3,3,3,3))
-dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey", fontsize_row=label_size(nrow(sampleDistMatrix)), fontsize_col=label_size(ncol(sampleDistMatrix)), show_rownames=label_size(nrow(sampleDistMatrix))>=4, show_colnames=label_size(ncol(sampleDistMatrix))>=4)
+dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey")
 save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.pdf")
 save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.png")
 save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.svg")
-#dend <- hclust(dist(mtcars, method = "euclidean"), method = "complete")
-#capture.output(cutree(dend, k = 3))
-#write.table(make_row_col_mat(sampleDistMatrix), file="differential_gene_expression/plots/Heatmaps_of_distances_index.tsv", row.#names=F, quote=F)
 ```
 
 ```{r heatmap_show, echo=FALSE, message=FALSE, warning=FALSE, out.width="90%", out.height="130%", fig.asp=1, dpi=1000, fig.align='center'}
@@ -1767,18 +1757,15 @@ for (file in contrast_files){
                     # Skip the heatmaps for CI tests
                     if (!isProvided(params$citest) & nrow(mat)>1){
                         dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/"))
-                        png(filename = paste("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png", sep=""), width = 2500, height = 3000, res = 300)
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+                        png(filename = paste("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png", sep=""), width = ifelse(nrow(mat)<=20), 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50)
+                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
-                        pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""))
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+                        pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""), width = ifelse(nrow(mat)<=20), 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
-                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""))
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""), width = ifelse(nrow(mat)<=20), 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
-                        dev.off()
-                        write.table(make_row_col_mat(mat), file=paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, "_index.tsv", sep=""), row.names=F, quote=F)
                     }
 
                     # Plotting pathway view only for kegg pathways
@@ -1854,18 +1841,15 @@ if (isProvided(params$path_genelist)){
     mat <- data.matrix(mat)
 
     if (nrow(mat)>1){
-        png(filename = paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png", sep=""), width = 2500, height = 3000, res = 300)
-        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
-        dev.off()
-        pdf(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf", sep=""))
-        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+        png(filename = paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png", sep=""), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100)
+        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
-        svg(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg", sep=""))
-        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+        pdf(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf", sep=""), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005)
+        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
-        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T, fontsize_row=label_size(nrow(mat)), fontsize_col=label_size(ncol(mat)), show_rownames=(label_size(nrow(mat))>=4), show_colnames=(label_size(ncol(mat))>=4))
+        svg(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg", sep=""), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+        pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
-        write.table(make_row_col_mat(mat), file=paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list_index.tsv", sep=""), row.names=F, quote=F)
     }
 }
 ```

From bbaa605c980a9ed9b29de475a9e5a790b2cff45f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 20 Feb 2023 15:10:27 +0100
Subject: [PATCH 015/124] bugfix

---
 assets/RNAseq_report.Rmd | 6 +++---
 1 file changed, 3 insertions(+), 3 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 87262a83..67c752a6 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1741,13 +1741,13 @@ for (file in contrast_files){
                     # Skip the heatmaps for CI tests
                     if (!isProvided(params$citest) & nrow(mat)>1){
                         dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/"))
-                        png(filename = paste("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png", sep=""), width = ifelse(nrow(mat)<=20), 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50)
+                        png(filename = paste("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png", sep=""), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50)
                         pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
-                        pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""), width = ifelse(nrow(mat)<=20), 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                        pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
                         pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
-                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""), width = ifelse(nrow(mat)<=20), 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
                         pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
                         dev.off()
                     }

From 7c4a4a1ddcc9b7a84a357d13cf8470ec7b8be9af Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 21 Feb 2023 11:12:50 +0100
Subject: [PATCH 016/124] Made software_versions optional

---
 CHANGELOG.md             |  2 ++
 assets/RNAseq_report.Rmd | 67 ++++++++++++++++++++++++++++++++--------
 modules/local/report.nf  |  3 +-
 nextflow.config          |  2 +-
 nextflow_schema.json     | 12 +++----
 workflows/rnadeseq.nf    |  5 ++-
 6 files changed, 67 insertions(+), 24 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 2ea1ab61..feb76e85 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -9,6 +9,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#200](https://github.com/qbic-pipelines/rnadeseq/pull/200) Made software_versions optional
+
 ### Fixed
 
 - [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 339077f3..ce451b48 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1968,12 +1968,15 @@ quote_string <- ifelse(
     paste0("The results for all work packages (no quote/offer given to describe them) can be found in this report.")
 )
 cat(quote_string)
+
+# Initiate var in case no software_versions were provided
+is_csv <- F
 ```
 
 Further support for this project will be restricted to the results presented in this report.
 For further analysis (e.g. the re-analysis of the dataset) we will generate a new quote containing cost estimates.
 
-```{r software_versions, bootstrap.show.message=FALSE, echo=FALSE, message=FALSE}
+```{r software_versions, bootstrap.show.message=FALSE, echo=FALSE, message=FALSE, eval=isProvided(params$path_software_versions)}
 # Import the .csv or .yml file output by the rna-seq Nextflow pipeline
 is_csv <- endsWith(params$path_software_versions, ".csv") || endsWith(params$path_software_versions, ".tsv")
 if (is_csv) {
@@ -2006,23 +2009,51 @@ if (is_csv) {
 
 # Computational methods
 
-## RNAseq data analysis
+```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
+
+# In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is obvious in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
+
+cat(paste0(
+"## RNAseq data analysis
+    
+The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(is_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
+"` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
+as.character(ifelse(is_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
+"` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
+Subsequently, adapter trimming was conducted with `Trim Galore ",
+as.character(ifelse(is_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
+"` [^4] [@krueger2012trim]. `STAR v",
+as.character(ifelse(is_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
+"` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
+The RNA-seq data quality control was performed with `RSeQC ",
+as.character(ifelse(is_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
+"` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
+quant_tool,
+" ",
+quant_version,
+"` ",
+quant_cite,
+"."
+))
 
-The Nextflow-based nf-core pipeline `rnaseq `r as.character(ifelse(is_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"]))`` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC `r as.character(ifelse(is_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"]))`` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
-Subsequently, adapter trimming was conducted with `Trim Galore `r as.character(ifelse(is_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"]))`` [^4] [@krueger2012trim]. `STAR v`r as.character(ifelse(is_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"]))`` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
-The RNA-seq data quality control was performed with `RSeQC `r as.character(ifelse(is_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"]))`` [@wang2012rseqc] and read quantification of the features (e.g. genes) with
-```{r quant_cite, echo=FALSE, results='asis', eval=T}
-cat(paste0("`", quant_tool, " ", quant_version, "` ", quant_cite, "."))
-```
-```{r MQC_cite, echo=FALSE, results='asis', eval=is_csv}
+```{r MQC_cite, echo=FALSE, results='asis', eval=is_csv&isProvided(params$path_software_versions)}
 cat(paste0("\nAn aggregation of the quality control for the RNAseq analysis was performed with `MultiQC ", as.character(version["MultiQC"]), "` [^2] [@ewels2016multiqc]."))
 ```
 
-## Differential gene expression analysis
+```{r software_versions_DE, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
 
-The differential expression analysis was performed by the use of the `rnadeseq `r as.character(params$revision)`` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `r quant_tool` were processed with the R package `DESeq2 v`r packageVersion("DESeq2")`` [@love2014differential].
+cat(paste0(
+"## Differential gene expression analysis
+
+The differential expression analysis was performed by the use of the `rnadeseq ",
+as.character(params$revision),
+"` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `",
+quant_tool, "` were processed with the R package `DESeq2 v",
+packageVersion("DESeq2"),
+"` [@love2014differential]."
+))
 
-```{r conditional_pathway2, echo=FALSE, results='asis', eval=params$pathway_analysis}
+```{r conditional_pathway, echo=FALSE, results='asis', eval=params$pathway_analysis&isProvided(params$path_software_versions)}
 version_gprofiler2 = packageVersion("gprofiler2")
 version_annotation = packageVersion("AnnotationDbi")
 name_species       = params$species_library
@@ -2042,7 +2073,17 @@ if (isProvided(params$custom_gmt)){
     database_string <- paste0(database_string, ".") # Add final dot to sentence
 }
 
-cat(paste0("For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2," `, `AnnotationDbi v", version_annotation, "` and `", name_species, " v", version_annotation, "` were employed. ", database_string))
+cat(paste0(
+"For pathway analysis, the R packages `gprofiler2 v",
+version_gprofiler2,
+" `, `AnnotationDbi v",
+version_annotation,
+"` and `",
+name_species,
+" v",
+version_annotation,
+"` were employed. ",
+database_string))
 ```
 
 Graphs were produced in RStudio with ``r R.version.string`` mainly using the R package `ggplot2 v`r packageVersion("ggplot2")`` . Final reports were produced using the R package `rmarkdown v`r packageVersion("rmarkdown") ``, with the `knitr v`r packageVersion("knitr")`` and `DT v`r packageVersion("DT")`` R packages.
diff --git a/modules/local/report.nf b/modules/local/report.nf
index a5395eeb..81a618b0 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -38,6 +38,7 @@ process REPORT {
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
 
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
+    def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : ''
 
     def citest_opt = params.citest ? "--citest TRUE" : ''
 
@@ -73,8 +74,8 @@ process REPORT {
         --keytype $params.keytype \
         --min_DEG_pathway $params.min_DEG_pathway \
         $quote_opt \
+        $software_versions_opt \
         --proj_summary $proj_summary \
-        --software_versions $software_versions \
         --revision $workflow.manifest.version \
         $citest_opt
 
diff --git a/nextflow.config b/nextflow.config
index f430db4c..72f9605f 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -16,7 +16,6 @@ params {
     input =  null
     model = null
     //* gtf = null
-    software_versions = null
 
     // DESeq2 args
     contrast_matrix = 'DEFAULT'
@@ -43,6 +42,7 @@ params {
     project_summary = null
     multiqc = 'NO_FILE4'
     quote = 'NO_FILE5'
+    software_versions = 'NO_FILE6'
 
     // Additional arg to indicate that a github test is run
     citest = false
diff --git a/nextflow_schema.json b/nextflow_schema.json
index ed2ad81d..5ba5c627 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -30,14 +30,9 @@
                 "gtf": {
                     "type": "string",
                     "description": "Not necessary if genome is set or input_type is featurecounts. GTF file to be used for DESeq if input is rsem or salmon."
-                },
-                "software_versions": {
-                    "type": "string",
-                    "default": "None",
-                    "description": "CSV/YML containing nf-core/rnaseq software versions."
                 }
             },
-            "required": ["input_type", "gene_counts", "input", "model", "software_versions"]
+            "required": ["input_type", "gene_counts", "input", "model"]
         },
         "deseq2_arguments": {
             "title": "DESeq2 arguments",
@@ -153,6 +148,11 @@
                 "quote": {
                     "type": "string",
                     "description": "Quote file to link in the report‚."
+                },
+                "software_versions": {
+                    "type": "string",
+                    "default": "None",
+                    "description": "CSV/YML containing nf-core/rnaseq software versions."
                 }
             }
         },
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
index d30c124d..07c634ff 100644
--- a/workflows/rnadeseq.nf
+++ b/workflows/rnadeseq.nf
@@ -13,7 +13,6 @@ WorkflowRnadeseq.initialise(params, log)
 // Check input path parameters to see if they exist
 def checkPathParamList = [
     params.input, params.model,
-    params.project_summary, params.software_versions
     ]
 for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } }
 
@@ -23,8 +22,6 @@ if (!(params.input_type in ["featurecounts", "salmon", "rsem"])) { exit 1, 'Wron
 if (params.gene_counts) { ch_counts_path = Channel.fromPath(params.gene_counts) } else { exit 1, 'Please provide input file/dir!' }
 if (params.input) { ch_metadata_file = Channel.fromPath(params.input) } else { exit 1, 'Please provide metadata file/samplesheet!' }
 if (params.model) { ch_model_file = Channel.fromPath(params.model) } else { exit 1, 'Please provide linear model file!' }
-if (params.project_summary) { ch_proj_summary_file = Channel.fromPath(params.project_summary) } else { exit 1, 'Please provide project summary file!' }
-if (params.software_versions) { ch_softwareversions_file = Channel.fromPath(params.software_versions) } else { exit 1, 'Please provide software versions file!' }
 
 // Create channel for genome parameter gtf (the other genome params are not files)
 if (params.input_type in ["rsem", "salmon"]) { ch_gtf = Channel.fromPath(params.gtf) } else { ch_gtf = Channel.fromPath("FALSE") }
@@ -37,6 +34,8 @@ ch_relevel = Channel.fromPath(params.relevel)
 ch_genes = Channel.fromPath(params.genelist)
 ch_multiqc_file = Channel.fromPath(params.multiqc)
 ch_custom_gmt = Channel.fromPath(params.custom_gmt)
+ch_proj_summary_file = Channel.fromPath(params.project_summary)
+ch_softwareversions_file = Channel.fromPath(params.software_versions)
 
 /*
 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

From cc3b89d78701cf608dbf144f7bb4b9608e5a0676 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 21 Feb 2023 11:20:10 +0100
Subject: [PATCH 017/124] Updated docs

---
 docs/usage.md | 66 +++++++++++++++++++++++++--------------------------
 1 file changed, 33 insertions(+), 33 deletions(-)

diff --git a/docs/usage.md b/docs/usage.md
index 92df57ed..9cdcf800 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -18,8 +18,6 @@
   - [`--input_type`](#--input_type)
   - [`--input`](#--input)
   - [`--model`](#--model)
-  - [`--project_summary`](#--project_summary)
-  - [`--software_versions`](#--software_versions)
 - [Contrasts](#contrasts)
   - [Default](#default)
   - [`--relevel`](#--relevel)
@@ -38,6 +36,8 @@
   - [`--run_pathway_analysis`](#--run_pathway_analysis)
   - [`--input_type`](#--input_type)
   - [`--multiqc`](#--multiqc)
+  - [`--project_summary`](#--project_summary)
+  - [`--software_versions`](#--software_versions)
   - [`--citest`](#--citest)
 - [Reference genome options](#reference-genome-options)
   - [`--genome`](#--genome)
@@ -216,37 +216,6 @@ Linear model function to calculate the contrasts (TXT). Variable names should be
 ~ condition_genotype + condition_treatment
 ```
 
-### `--project_summary`
-
-Project summary as downloaded from the portal: User database portlet, Projects tab, select your project and "Download Project Information". Please check first that this information is correct in the project Browser.
-
-### `--software_versions`
-
-Path to the `Software_versions.csv/.yml` file that is generated by the nf-core/rnaseq pipeline.
-
-The CSV file should be tab-separated:
-
-```bash
-nf-core/rnaseq	v1.4dev
-Nextflow	v19.01.0
-FastQC	v0.11.8
-Cutadapt	v2.1
-Trim Galore!	v0.5.0
-STAR	vSTAR_2.6.1d
-```
-
-Alternatively, the YML file should look like this:
-
-```bash
-BEDTOOLS_GENOMECOV:
-  bedtools: 2.30.0
-CUSTOM_DUMPSOFTWAREVERSIONS:
-  python: 3.10.6
-  yaml: '6.0'
-CUSTOM_GETCHROMSIZES:
-  custom: 1.15.1
-```
-
 ## Contrasts
 
 There are three different parameters that can be used to define contrasts, which are explained in the following sections. One or multiple contrast input files can be provided, if multiple are provided, the contrasts in the multiple files will be added to the report.
@@ -357,6 +326,37 @@ This tells the pipeline which type of input dataset is provided. Must be one of
 
 Path to the MultiQC zipped folder containing the MultiQC plots generated by the RNAseq pipeline, the MultiQC html report, and the MultiQC raw data; optional.
 
+### `--project_summary`
+
+Project summary as downloaded from the portal: User database portlet, Projects tab, select your project and "Download Project Information". Please check first that this information is correct in the project Browser.
+
+### `--software_versions`
+
+Path to the `Software_versions.csv/.yml` file that is generated by the nf-core/rnaseq pipeline.
+
+The CSV file should be tab-separated:
+
+```bash
+nf-core/rnaseq	v1.4dev
+Nextflow	v19.01.0
+FastQC	v0.11.8
+Cutadapt	v2.1
+Trim Galore!	v0.5.0
+STAR	vSTAR_2.6.1d
+```
+
+Alternatively, the YML file should look like this:
+
+```bash
+BEDTOOLS_GENOMECOV:
+  bedtools: 2.30.0
+CUSTOM_DUMPSOFTWAREVERSIONS:
+  python: 3.10.6
+  yaml: '6.0'
+CUSTOM_GETCHROMSIZES:
+  custom: 1.15.1
+```
+
 ### `--citest`
 
 Developer param, tells the pipeline it is run on a github server which affects how output files are saved.

From ac55c9d23149bafb0a758702fb464316eaeb1b97 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 1 Mar 2023 08:13:30 +0100
Subject: [PATCH 018/124] Renamed variable

---
 assets/RNAseq_report.Rmd | 22 +++++++++++-----------
 1 file changed, 11 insertions(+), 11 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index ce451b48..0260055e 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1970,7 +1970,7 @@ quote_string <- ifelse(
 cat(quote_string)
 
 # Initiate var in case no software_versions were provided
-is_csv <- F
+software_versions_csv <- F
 ```
 
 Further support for this project will be restricted to the results presented in this report.
@@ -1978,21 +1978,21 @@ For further analysis (e.g. the re-analysis of the dataset) we will generate a ne
 
 ```{r software_versions, bootstrap.show.message=FALSE, echo=FALSE, message=FALSE, eval=isProvided(params$path_software_versions)}
 # Import the .csv or .yml file output by the rna-seq Nextflow pipeline
-is_csv <- endsWith(params$path_software_versions, ".csv") || endsWith(params$path_software_versions, ".tsv")
-if (is_csv) {
+software_versions_csv <- endsWith(params$path_software_versions, ".csv") || endsWith(params$path_software_versions, ".tsv")
+if (software_versions_csv) {
     version_csv <- read.csv(file = params$path_software_versions, sep="\t", header = FALSE)
     version <- setNames(as.list(version_csv$V2), version_csv$V1)
 } else {
     version <- read_yaml(params$path_software_versions)
 }
 
-if (is_csv) {
+if (software_versions_csv) {
     if (params$input_type == "featurecounts") {
         quant_tool <- "featureCounts"
         quant_version <- as.character(version["featureCounts"])
         quant_cite <- "[@liao2014featurecounts]"
     }
-} else if (!(is_csv)) {
+} else if (!(software_versions_csv)) {
     if (params$input_type == "rsem") {
         quant_tool <- "RSEM"
         quant_version <- as.character(version["RSEM_CALCULATEEXPRESSION"][[1]]["rsem"])
@@ -2016,17 +2016,17 @@ if (is_csv) {
 cat(paste0(
 "## RNAseq data analysis
     
-The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(is_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
+The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(software_versions_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
 "` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
-as.character(ifelse(is_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
+as.character(ifelse(software_versions_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
 "` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
 Subsequently, adapter trimming was conducted with `Trim Galore ",
-as.character(ifelse(is_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
+as.character(ifelse(software_versions_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
 "` [^4] [@krueger2012trim]. `STAR v",
-as.character(ifelse(is_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
+as.character(ifelse(software_versions_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
 "` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
 The RNA-seq data quality control was performed with `RSeQC ",
-as.character(ifelse(is_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
+as.character(ifelse(software_versions_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
 "` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
 quant_tool,
 " ",
@@ -2036,7 +2036,7 @@ quant_cite,
 "."
 ))
 
-```{r MQC_cite, echo=FALSE, results='asis', eval=is_csv&isProvided(params$path_software_versions)}
+```{r MQC_cite, echo=FALSE, results='asis', eval=software_versions_csv&isProvided(params$path_software_versions)}
 cat(paste0("\nAn aggregation of the quality control for the RNAseq analysis was performed with `MultiQC ", as.character(version["MultiQC"]), "` [^2] [@ewels2016multiqc]."))
 ```
 

From fbfadb6ae2b077e6897a3498786def139f9d5fee Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 1 Mar 2023 14:18:57 +0100
Subject: [PATCH 019/124] Updated some 5sums, set a logFC for test_full in the
 hopes it will run faster on github now

---
 conf/test_batcheffect.config |  1 -
 conf/test_full.config        |  1 +
 tests/test.yml               | 10 +++++-----
 tests/test_star_rsem.yml     | 16 ++++++++--------
 tests/test_star_salmon.yml   |  8 ++++----
 5 files changed, 18 insertions(+), 18 deletions(-)

diff --git a/conf/test_batcheffect.config b/conf/test_batcheffect.config
index 2b7d43a2..0f8293aa 100644
--- a/conf/test_batcheffect.config
+++ b/conf/test_batcheffect.config
@@ -22,7 +22,6 @@ params {
     gene_counts = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/merged_gene_counts.txt'
     model = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/design_batcheffect.txt'
     genelist = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/requested_genes.txt'
-    logFC_threshold = 1
     //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml'
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
     software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv'
diff --git a/conf/test_full.config b/conf/test_full.config
index 66cc60f9..30f05b24 100644
--- a/conf/test_full.config
+++ b/conf/test_full.config
@@ -27,6 +27,7 @@ params {
     software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv'
     multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip'
     run_pathway_analysis = true
+    logFC_threshold = 1
     genome = 'GRCm38'
     quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf'
 }
diff --git a/tests/test.yml b/tests/test.yml
index 78d5c602..d6e6a0bd 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -63,11 +63,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: 53d777593af962a4b590cef2e076780c
+      md5sum: e6a4b7d7ee1d993da4f8fd1359f38bbe
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 75eba6f4796f03efc3321f2427afe18e
+      md5sum: 28f1f81ea5c50eaf77e5733845c25393
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 15f0769b63d766b056f7298ab6a3a2a8
+      md5sum: 0daf33fba37e4a03752858c1a8d60b74
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf
@@ -79,9 +79,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: fa0b5b0772d026ff5a6b5ccffb05eb14
+      md5sum: 4ceb132e9d7f8a5c40894b4e1459b2a0
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: 2e896e28c5a064767dda6668b292e064
+      md5sum: 1939189eee32b27500f37414f716fef2
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 6664bb6f..d02abd0e 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -6,11 +6,11 @@
     - path: results_test/differential_gene_expression/metadata/star_rsem_files.txt
       md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
-      md5sum: 216106ef902281c5d7c72715583e384c
+      md5sum: 60aff9f171a58cd2c9db9e60eda3a506
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
       md5sum: 7a4db95674a9c04aaafaef339edcfcc7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_MCF7_vs_GM12878.tsv
-      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
+      md5sum: b7e914acf8a0d379e0ace343cfd1a69b
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: f1825ed59878115a279db7151b0ed0a1
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/CXCL2_ENSG00000081041.pdf
@@ -88,11 +88,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 569d992498f2feb6eab5186b272510bf
+      md5sum: 42c6e198d0d3a7a28e73ce39a6e509c9
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: d93e805f242f9f16ad99a94602123468
+      md5sum: dbc9fb3692f193d660f549fa8dcf2ea4
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 51266349ce9a9bfc62ea4ea149dcc10f
+      md5sum: a4e9e28b094474383ce5f6d208871562
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
@@ -108,9 +108,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/KEGG_pathways/hsa01040.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/KEGG_pathways/hsa01040.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv
-      md5sum: 46ce910756ee4dc8a96f310838d5d398
+      md5sum: c5591023437dabcf0279ebc4f2a12771
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: d93e805f242f9f16ad99a94602123468
+      md5sum: dbc9fb3692f193d660f549fa8dcf2ea4
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv
-      md5sum: 7d1dc63865de1c1768cb536fedc53519
+      md5sum: 96bd281d46bb3708c840f222f9559ab0
     - path: results_test/RNAseq_report.html
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index 8b424779..ed0ffe17 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -70,13 +70,13 @@
     - path: results_test/differential_gene_expression/plots/PCA_plot.png
     - path: results_test/differential_gene_expression/plots/PCA_plot.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 68b329da9893e34099c7d8ad5cb9c940
+      md5sum: 8c6ed54d7faf0beb6d41afa9019c8857
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv
-      md5sum: 45af78cf5aec148366d9a25acdf71f13
+      md5sum: 23ac7b3845e539fa5748fe58a920fd89
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: b118b02cf5ce4716b7bcac06cde37c83
+      md5sum: 1b4ae63b60287794f8eb4d1fcaf04fa3
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv
-      md5sum: 40b83ea71cef466f02da524ce4b16701
+      md5sum: eb7e714867d0afb0f851b09ebf2c5a88
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/RNAseq_report.html

From 66a6125703587514dd34b974620ea52dee50ab37 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 1 Mar 2023 16:10:49 +0100
Subject: [PATCH 020/124] updated 5sums for test_full after adding logfc

---
 tests/test_full.yml | 16 ++++++++--------
 1 file changed, 8 insertions(+), 8 deletions(-)

diff --git a/tests/test_full.yml b/tests/test_full.yml
index bbf5bc70..d2a6baee 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -4,11 +4,11 @@
     - test_full
   files:
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_genotype_WT_vs_KO.tsv
-      md5sum: b19e34f880a7cb14e031a00d6c8f1b74
+      md5sum: 93fd663cb976c102144e1ffb34d88e53
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_Treated_vs_Control.tsv
-      md5sum: 05244958729524ea28c903caf1161e77
+      md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
-      md5sum: dc900cdde288863cb51fac6111fac761
+      md5sum: 81d3437334849b3b4de4de6729a3d54d
     - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
@@ -63,11 +63,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: d55f1b765fe7105bfb88f8e68048d68c
+      md5sum: e6a4b7d7ee1d993da4f8fd1359f38bbe
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 71e3e0cd3fbd529d5648802487849bc5
+      md5sum: 28f1f81ea5c50eaf77e5733845c25393
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 12c145521c9fe149f21beb5c2c14c094
+      md5sum: 0daf33fba37e4a03752858c1a8d60b74
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf
@@ -76,9 +76,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 3316eebfe0f6916c62afa4e9ee9eeb91
+      md5sum: 4ceb132e9d7f8a5c40894b4e1459b2a0
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: 616e96c92b53aac70a410b31ab7470d0
+      md5sum: 1939189eee32b27500f37414f716fef2
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf

From 2ea47a4b340f1e9005f65e27cb89e3ce3a681c13 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 2 Mar 2023 09:02:22 +0100
Subject: [PATCH 021/124] Updated md5sums

---
 tests/test_star_rsem.yml | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index d02abd0e..c6917d0c 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -6,11 +6,11 @@
     - path: results_test/differential_gene_expression/metadata/star_rsem_files.txt
       md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
-      md5sum: 60aff9f171a58cd2c9db9e60eda3a506
+      md5sum: 216106ef902281c5d7c72715583e384c
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
       md5sum: 7a4db95674a9c04aaafaef339edcfcc7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_MCF7_vs_GM12878.tsv
-      md5sum: b7e914acf8a0d379e0ace343cfd1a69b
+      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: f1825ed59878115a279db7151b0ed0a1
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/CXCL2_ENSG00000081041.pdf

From 45626719071e8f7634a1785783baefac22082304 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 2 Mar 2023 11:03:50 +0100
Subject: [PATCH 022/124] commented out two md5sums as they constantly change
 even without environment change

---
 tests/test_star_rsem.yml | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index c6917d0c..8732a767 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -6,11 +6,11 @@
     - path: results_test/differential_gene_expression/metadata/star_rsem_files.txt
       md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
-      md5sum: 216106ef902281c5d7c72715583e384c
+#      md5sum: 216106ef902281c5d7c72715583e384c
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
       md5sum: 7a4db95674a9c04aaafaef339edcfcc7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_MCF7_vs_GM12878.tsv
-      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
+#      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: f1825ed59878115a279db7151b0ed0a1
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/CXCL2_ENSG00000081041.pdf

From d8a39ffe7e18c513f9e9f34e8333ebc2e00bf7cf Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 2 Mar 2023 12:35:16 +0100
Subject: [PATCH 023/124] prettier

---
 tests/test_star_rsem.yml | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 8732a767..933fded7 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -6,11 +6,11 @@
     - path: results_test/differential_gene_expression/metadata/star_rsem_files.txt
       md5sum: ad72a01afb6a17864acac149c636df98
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_H1_vs_GM12878.tsv
-#      md5sum: 216106ef902281c5d7c72715583e384c
+    #      md5sum: 216106ef902281c5d7c72715583e384c
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_K562_vs_GM12878.tsv
       md5sum: 7a4db95674a9c04aaafaef339edcfcc7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_cellline_MCF7_vs_GM12878.tsv
-#      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
+    #      md5sum: ea4b16a379f09fa44c7c541ccab64ce6
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: f1825ed59878115a279db7151b0ed0a1
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/CXCL2_ENSG00000081041.pdf

From 1d013ec2359d79960be815f9d99c04c4a8bbd3a6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 6 Mar 2023 15:22:43 +0100
Subject: [PATCH 024/124] still testing how the background list works

---
 assets/RNAseq_report.Rmd |  7 +++++--
 bin/Execute_report.R     |  2 ++
 modules/local/report.nf  |  3 +++
 nextflow.config          |  3 ++-
 nextflow_schema.json     | 12 ++++++++----
 workflows/rnadeseq.nf    |  4 +++-
 6 files changed, 23 insertions(+), 8 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 67c752a6..7529eafb 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -40,6 +40,7 @@ params:
     keytype: ''
     custom_gmt: ''
     min_DEG_pathway: ''
+    custom_background: ''
 
     #Additional args for the report
     path_proj_summary: ''
@@ -1648,7 +1649,8 @@ for (file in contrast_files){
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
-            user_threshold=params$pval_threshold)
+            user_threshold=params$pval_threshold
+)
 
     # Make data frame of gost result
     pathway_gostres <- gostres$result
@@ -1908,7 +1910,8 @@ if (length(q_list) > 0) {
                 significant=T,
                 correction_method="fdr",
                 sources=datasources,
-                user_threshold=params$pval_threshold)
+                user_threshold=params$pval_threshold
+)
     path_enrich <- as.data.frame(gostres$result)
     if (nrow(path_enrich) > 0){
         pg2 <- gostplot(gostres, capped=T, interactive=T)
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index 91125536..678dec22 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -31,6 +31,7 @@ option_list = list(
     make_option(c("-i", "--species_library"), type="character", default=NULL, help="Library name. Example format: org.At.tair.db", metavar="character"),
     make_option(c("-u", "--keytype"), type="character", default=NULL, help="Keytype. Example format: TAIR (varies greatly depending on library!)", metavar="character"),
     make_option("--custom_gmt", type="character", default=NULL, help="Path to custom GMT file to query during pathway analysis.", metavar="character"),
+    make_option("--custom_background", type="character", default=NULL, help="Path to background TXT file of gene names/IDs to consider during pathway analysis.", metavar="character"),
     make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"),
 
     make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"),
@@ -76,6 +77,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 organism = opt$organism,
                                 species_library = opt$species_library,
                                 custom_gmt = opt$custom_gmt,
+                                custom_background = opt$custom_background,
                                 keytype = opt$keytype,
                                 min_DEG_pathway = opt$min_DEG_pathway,
 
diff --git a/modules/local/report.nf b/modules/local/report.nf
index a5395eeb..d14baf7f 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -18,6 +18,7 @@ process REPORT {
     path software_versions
     path multiqc
     path custom_gmt
+    path custom_background
 
     output:
     path "*.zip"
@@ -36,6 +37,7 @@ process REPORT {
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
+    def custom_background_opt = custom_background.name != 'NO_FILE6' ? "--custom_background $custom_background" : ''
 
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
 
@@ -68,6 +70,7 @@ process REPORT {
         $round_DE_opt \
         $pathway_opt \
         $custom_gmt_opt \
+        $custom_background_opt \
         --organism $params.organism \
         --species_library $params.species_library \
         --keytype $params.keytype \
diff --git a/nextflow.config b/nextflow.config
index f430db4c..26cde721 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -35,9 +35,10 @@ params {
     //* organism = null
     //* species_library = null
     //* keytype = null
-    custom_gmt = 'NO_FILE3'
     run_pathway_analysis = false
     min_DEG_pathway = 1
+    custom_gmt = 'NO_FILE3'
+    custom_background = 'NO_FILE6'
 
     // Additional args for the report
     project_summary = null
diff --git a/nextflow_schema.json b/nextflow_schema.json
index ed2ad81d..46be9be3 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -120,15 +120,19 @@
                     "type": "string",
                     "description": "Not necessary if genome is set. Which keytype to use for pathway analysis, e.g. ENSEMBL, not necessary if --run_pathway_analysis = false."
                 },
-                "custom_gmt": {
-                    "type": "string",
-                    "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true."
-                },
                 "run_pathway_analysis": {
                     "type": "boolean",
                     "default": false,
                     "description": "Turn on this flag if you wish to run pathway analysis."
                 },
+                "custom_gmt": {
+                    "type": "string",
+                    "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true."
+                },
+                "custom_background": {
+                    "type": "string",
+                    "description": "Path to TXT file containing a background list of genes for pathway analysis (one gene name/ID per line)."
+                },
                 "min_DEG_pathway": {
                     "type": "integer",
                     "default": 1,
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
index d30c124d..736bf1fc 100644
--- a/workflows/rnadeseq.nf
+++ b/workflows/rnadeseq.nf
@@ -37,6 +37,7 @@ ch_relevel = Channel.fromPath(params.relevel)
 ch_genes = Channel.fromPath(params.genelist)
 ch_multiqc_file = Channel.fromPath(params.multiqc)
 ch_custom_gmt = Channel.fromPath(params.custom_gmt)
+ch_custom_background = Channel.fromPath(params.custom_background)
 
 /*
 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
@@ -99,7 +100,8 @@ workflow RNADESEQ {
         ch_proj_summary_file,
         ch_softwareversions_file,
         ch_multiqc_file,
-        ch_custom_gmt
+        ch_custom_gmt,
+        ch_custom_background
     )
 
     //TODO: Enable this:

From c9482ad792cc0d296e3e94d7e19b15a40bddc8ba Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 8 Mar 2023 08:16:08 +0100
Subject: [PATCH 025/124] readded custom_bg to gost()

---
 assets/RNAseq_report.Rmd | 6 ++++--
 1 file changed, 4 insertions(+), 2 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 7529eafb..48d16be1 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1649,7 +1649,8 @@ for (file in contrast_files){
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
-            user_threshold=params$pval_threshold
+            user_threshold=params$pval_threshold,
+            custom_bg=rownames(counts(cds))[notAllZero]
 )
 
     # Make data frame of gost result
@@ -1910,7 +1911,8 @@ if (length(q_list) > 0) {
                 significant=T,
                 correction_method="fdr",
                 sources=datasources,
-                user_threshold=params$pval_threshold
+                user_threshold=params$pval_threshold,
+                custom_bg=rownames(counts(cds))[notAllZero]
 )
     path_enrich <- as.data.frame(gostres$result)
     if (nrow(path_enrich) > 0){

From 26405dfa8c141aaf91a60a2191b2247c3b182a69 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 08:23:46 +0100
Subject: [PATCH 026/124] Finalized the background list logic

---
 assets/RNAseq_report.Rmd | 94 +++++++++++++++++++++++++++++++++-------
 bin/Execute_report.R     |  6 +--
 environment.yml          |  1 +
 modules/local/report.nf  |  5 +--
 nextflow.config          |  2 +-
 nextflow_schema.json     |  7 +--
 workflows/rnadeseq.nf    |  4 +-
 7 files changed, 91 insertions(+), 28 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index ed4b5b90..0cf78ce8 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -38,9 +38,9 @@ params:
     organism: ''
     species_library: ''
     keytype: ''
-    custom_gmt: ''
     min_DEG_pathway: ''
-    custom_background: ''
+    custom_gmt: ''
+    background_list: ''
 
     #Additional args for the report
     path_proj_summary: ''
@@ -1530,7 +1530,8 @@ invisible(lapply(c(
 "reshape2",
 "pathview",
 "AnnotationDbi",
-"optparse"
+"optparse",
+"ggvenn"
 ), library, character.only=T))
 
 # Create output directory
@@ -1643,24 +1644,57 @@ for (file in contrast_files){
     q = as.character(DE_genes$Ensembl_ID)
 
     # gost query
-    gostres <- gost(query=q,
+    if (params$background_list) {
+        gostres <- gost(query=q,
             organism=gost_id,
             significant=TRUE,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
             user_threshold=params$pval_threshold,
-            custom_bg=rownames(counts(cds))[notAllZero]
-)
+            custom_bg=rownames(counts(cds))[notAllZero],
+            domain_scope="custom_annotated"
+        )
+        
+        pathway_gostres <- gostres$result
+        pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),])
+        pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),]
+        
+        # Also run without bg for Venn diagrams
+        gostres_nobg <- gost(query=q,
+            organism=gost_id,
+            significant=TRUE,
+            correction_method="fdr",
+            sources=datasources,
+            evcodes=TRUE,
+            user_threshold=params$pval_threshold,
+            domain_scope="annotated"
+        )
+        pathway_gostres_nobg <- gostres_nobg$result
+        pathway_gostres_nobg <- as.data.frame(pathway_gostres_nobg[which(pathway_gostres_nobg$significant==TRUE),])
+        pathway_gostres_nobg <- pathway_gostres_nobg[which(pathway_gostres_nobg$intersection_size>=min_DEG_pathway),]
+
+    } else {
+        gostres <- gost(query=q,
+            organism=gost_id,
+            significant=TRUE,
+            correction_method="fdr",
+            sources=datasources,
+            evcodes=TRUE,
+            user_threshold=params$pval_threshold,
+            domain_scope="annotated"
+        )
+    }
+
 
     # Make data frame of gost result
     pathway_gostres <- gostres$result
+
     # Select only significantly enriched pathways (according to adjusted p-value)
     pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),])
 
     # Select only pathways with a min. number of DEG
     pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),]
-
     gostres$result <- pathway_gostres
 
     # Plot pathways if there were any
@@ -1680,7 +1714,23 @@ for (file in contrast_files){
         ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.svg"),
             device="svg",
             height=10, width=15, units="cm", dpi=300, limitsize=F)
+
+        # If background list was used for main PA analysis, plot Venn diagram of the results and those from the additional PA without background list
+        if (params$background_list) {
+            pathway_gostres_nobg <- gostres_nobg$result
+            pathway_gostres_nobg <- as.data.frame(pathway_gostres_nobg[which(pathway_gostres_nobg$significant==TRUE),])
+            pathway_gostres_nobg <- pathway_gostres_nobg[which(pathway_gostres_nobg$intersection_size>=min_DEG_pathway),]
+            VennInput <- list(
+                background=pathway_gostres$term_name,
+                no_background=pathway_gostres_nobg$term_name
+            )
+            pv <- ggvenn::ggvenn(VennInput)
+            ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.pdf"),pv, device="pdf", width=10, height=15)
+            ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.png"),pv, device="png", width=10, height=15)
+            ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.svg"),pv, device="svg", width=10, height=15)
+        }
     }
+
     # Remove parents column to be able to save the table in tsv format
     pathway_gostres_table <- pathway_gostres
     pathway_gostres_table$parents <- NULL
@@ -1906,14 +1956,28 @@ names(q_list) <- q_names
 if (length(q_list) > 0) {
 
     #gost query
-    gostres <- gost(query=q_list,
-                organism=gost_id,
-                significant=T,
-                correction_method="fdr",
-                sources=datasources,
-                user_threshold=params$pval_threshold,
-                custom_bg=rownames(counts(cds))[notAllZero]
-)
+    
+    if (params$background_list) {
+        gostres <- gost(query=q_list,
+                    organism=gost_id,
+                    significant=T,
+                    correction_method="fdr",
+                    sources=datasources,
+                    user_threshold=params$pval_threshold,
+                    custom_bg=rownames(counts(cds))[notAllZero],
+                    domain_scope="custom_annotated"
+        )
+    } else {
+        gostres <- gost(query=q_list,
+                    organism=gost_id,
+                    significant=T,
+                    correction_method="fdr",
+                    sources=datasources,
+                    user_threshold=params$pval_threshold,
+                    domain_scope="annotated"
+        )
+    }
+
     path_enrich <- as.data.frame(gostres$result)
     if (nrow(path_enrich) > 0){
         pg2 <- gostplot(gostres, capped=T, interactive=T)
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index 678dec22..4246cea0 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -26,12 +26,12 @@ option_list = list(
     make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
     make_option(c("-n", "--nsub_genes"), type="integer", default=NULL, help="Subset number of genes for vst."),
 
-    make_option(c("-a", "--pathway_analysis"), action="store_true", default=FALSE, help="Pathway analysis."),
+    make_option(c("-a", "--pathway_analysis"), action="store_true", default=FALSE, help="Whether to run pathway analysis."),
     make_option(c("-g", "--organism"), type="character", default=NULL, help="Organism, e.g. Hsapiens."),
     make_option(c("-i", "--species_library"), type="character", default=NULL, help="Library name. Example format: org.At.tair.db", metavar="character"),
     make_option(c("-u", "--keytype"), type="character", default=NULL, help="Keytype. Example format: TAIR (varies greatly depending on library!)", metavar="character"),
     make_option("--custom_gmt", type="character", default=NULL, help="Path to custom GMT file to query during pathway analysis.", metavar="character"),
-    make_option("--custom_background", type="character", default=NULL, help="Path to background TXT file of gene names/IDs to consider during pathway analysis.", metavar="character"),
+    make_option("--background_list", action="store_true", default=TRUE, help="Whether to use a background list for pathway analysis; if true, will only consider expressed genes (i.e. mean counts > 0) for PA."),
     make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"),
 
     make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"),
@@ -77,7 +77,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 organism = opt$organism,
                                 species_library = opt$species_library,
                                 custom_gmt = opt$custom_gmt,
-                                custom_background = opt$custom_background,
+                                background_list = opt$background_list,
                                 keytype = opt$keytype,
                                 min_DEG_pathway = opt$min_DEG_pathway,
 
diff --git a/environment.yml b/environment.yml
index 89519875..fe104828 100644
--- a/environment.yml
+++ b/environment.yml
@@ -31,6 +31,7 @@ dependencies:
   - conda-forge::r-formattable=0.2.1
   - conda-forge::r-ggplot2=3.3.5
   - conda-forge::r-ggrepel=0.9.2
+  - conda-forge::r-ggvenn=0.1.9
   - conda-forge::r-gplots=3.1.1
   - conda-forge::r-gprofiler2=0.2.1
   - conda-forge::r-heatmaply=1.3.0
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 64138db3..e7c70866 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -18,7 +18,6 @@ process REPORT {
     path software_versions
     path multiqc
     path custom_gmt
-    path custom_background
 
     output:
     path "*.zip"
@@ -37,7 +36,7 @@ process REPORT {
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
-    def custom_background_opt = custom_background.name != 'NO_FILE6' ? "--custom_background $custom_background" : ''
+    def background_list_opt = $params.background_list ? "--background_list TRUE" : "--background_list FALSE"
 
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
     def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : ''
@@ -71,7 +70,7 @@ process REPORT {
         $round_DE_opt \
         $pathway_opt \
         $custom_gmt_opt \
-        $custom_background_opt \
+        $background_list_opt \
         --organism $params.organism \
         --species_library $params.species_library \
         --keytype $params.keytype \
diff --git a/nextflow.config b/nextflow.config
index f66ea905..476d21cb 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -37,7 +37,7 @@ params {
     run_pathway_analysis = false
     min_DEG_pathway = 1
     custom_gmt = 'NO_FILE3'
-    custom_background = 'NO_FILE6'
+    background_list = true
 
     // Additional args for the report
     project_summary = null
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 6bfe9400..2f5ae149 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -124,9 +124,10 @@
                     "type": "string",
                     "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true."
                 },
-                "custom_background": {
-                    "type": "string",
-                    "description": "Path to TXT file containing a background list of genes for pathway analysis (one gene name/ID per line)."
+                "background_list": {
+                    "type": "boolean",
+                    "default": true,
+                    "description": "Whether to limit pathway analysis to a background list of genes (i.e. those genes expressed in at least one sample), default = true."
                 },
                 "min_DEG_pathway": {
                     "type": "integer",
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
index 58bb8cd0..07c634ff 100644
--- a/workflows/rnadeseq.nf
+++ b/workflows/rnadeseq.nf
@@ -34,7 +34,6 @@ ch_relevel = Channel.fromPath(params.relevel)
 ch_genes = Channel.fromPath(params.genelist)
 ch_multiqc_file = Channel.fromPath(params.multiqc)
 ch_custom_gmt = Channel.fromPath(params.custom_gmt)
-ch_custom_background = Channel.fromPath(params.custom_background)
 ch_proj_summary_file = Channel.fromPath(params.project_summary)
 ch_softwareversions_file = Channel.fromPath(params.software_versions)
 
@@ -99,8 +98,7 @@ workflow RNADESEQ {
         ch_proj_summary_file,
         ch_softwareversions_file,
         ch_multiqc_file,
-        ch_custom_gmt,
-        ch_custom_background
+        ch_custom_gmt
     )
 
     //TODO: Enable this:

From cd8380ba6251a8cb11f8c37e0357ef351c766048 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 08:57:17 +0100
Subject: [PATCH 027/124] fixed error, added Venn diagrams to one test config

---
 modules/local/report.nf    | 2 +-
 tests/test_batcheffect.yml | 2 +-
 tests/test_full.yml        | 3 +++
 3 files changed, 5 insertions(+), 2 deletions(-)

diff --git a/modules/local/report.nf b/modules/local/report.nf
index e7c70866..e59e45ea 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -36,7 +36,7 @@ process REPORT {
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
-    def background_list_opt = $params.background_list ? "--background_list TRUE" : "--background_list FALSE"
+    def background_list_opt = params.background_list ? "--background_list TRUE" : "--background_list FALSE"
 
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
     def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : ''
diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml
index afac9599..7afa1153 100644
--- a/tests/test_batcheffect.yml
+++ b/tests/test_batcheffect.yml
@@ -1,5 +1,5 @@
 - name: Run test profile with batcheffect correction
-  command: nextflow run main.nf -profile test_batcheffect,docker --citest true --round_DE 3
+  command: nextflow run main.nf -profile test_batcheffect,docker --citest true --round_DE 3 --background_list false
   tags:
     - test_batcheffect
   files:
diff --git a/tests/test_full.yml b/tests/test_full.yml
index d2a6baee..6079a602 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -68,6 +68,9 @@
       md5sum: 28f1f81ea5c50eaf77e5733845c25393
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
       md5sum: 0daf33fba37e4a03752858c1a8d60b74
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf

From 039c9a1b6298ef8ae70ac2258420e4a39797848f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 10:08:28 +0100
Subject: [PATCH 028/124] updated changelog and md5sums

---
 CHANGELOG.md               |  1 +
 tests/test.yml             | 10 +++++-----
 tests/test_custom_gmt.yml  |  4 ++--
 tests/test_full.yml        | 10 +++++-----
 tests/test_star_rsem.yml   | 14 ++++++--------
 tests/test_star_salmon.yml | 10 ++--------
 6 files changed, 21 insertions(+), 28 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 467cf46b..26b67dea 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
+- [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
 - [#197](https://github.com/qbic-pipelines/rnadeseq/pull/197) Added gprofiler version string to report
 - [#196](https://github.com/qbic-pipelines/rnadeseq/pull/196) Added optional `--quote` parameter
 
diff --git a/tests/test.yml b/tests/test.yml
index d6e6a0bd..64ca823b 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -63,11 +63,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: e6a4b7d7ee1d993da4f8fd1359f38bbe
+      md5sum: 237e9630d2998badcc69b1e193d5e1ab
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 28f1f81ea5c50eaf77e5733845c25393
+      md5sum: 7ee7cd938140217a863774c3689ff1af
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 0daf33fba37e4a03752858c1a8d60b74
+      md5sum: 18873d39d839477337312cce0ec63398
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf
@@ -79,9 +79,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 4ceb132e9d7f8a5c40894b4e1459b2a0
+      md5sum: 9318d57b86ba653f6183e61840a7943b
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: 1939189eee32b27500f37414f716fef2
+      md5sum: d5632f9240116f42de59f05b7e822a29
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml
index a306e24c..5a9dec79 100644
--- a/tests/test_custom_gmt.yml
+++ b/tests/test_custom_gmt.yml
@@ -63,8 +63,8 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: 058c5c9e251d08ec087475aa2b55cdff
+      md5sum: eae154d9c7a526cc8c3fcd1a586fa0ed
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 8606aafe70ae5c6ce9bc4ce4b414ca4f
+      md5sum: bd2cbbf85c3e241743e60b9b8263a31c
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
     - path: results_test/RNAseq_report.html
diff --git a/tests/test_full.yml b/tests/test_full.yml
index 6079a602..02cd336a 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -63,11 +63,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: e6a4b7d7ee1d993da4f8fd1359f38bbe
+      md5sum: 237e9630d2998badcc69b1e193d5e1ab
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 28f1f81ea5c50eaf77e5733845c25393
+      md5sum: 7ee7cd938140217a863774c3689ff1af
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 0daf33fba37e4a03752858c1a8d60b74
+      md5sum: 18873d39d839477337312cce0ec63398
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg
@@ -79,9 +79,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 4ceb132e9d7f8a5c40894b4e1459b2a0
+      md5sum: 9318d57b86ba653f6183e61840a7943b
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: 1939189eee32b27500f37414f716fef2
+      md5sum: d5632f9240116f42de59f05b7e822a29
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 933fded7..377b99a8 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -88,11 +88,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 42c6e198d0d3a7a28e73ce39a6e509c9
+      md5sum: 7f71760d73cab3c0d76a59c5ecdd78c0
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: dbc9fb3692f193d660f549fa8dcf2ea4
+      md5sum: 2b2ae0c1d4b4de442ced8d37193f61d4
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: a4e9e28b094474383ce5f6d208871562
+      md5sum: a5422d271a102dff6c14ecfa46abdeab
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
@@ -105,12 +105,10 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/KEGG_pathways/hsa01040.png
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/KEGG_pathways/hsa01040.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv
-      md5sum: c5591023437dabcf0279ebc4f2a12771
+      md5sum: 210529cfe4281b056e5bd59550757295
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: dbc9fb3692f193d660f549fa8dcf2ea4
+      md5sum: 2b2ae0c1d4b4de442ced8d37193f61d4
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv
-      md5sum: 96bd281d46bb3708c840f222f9559ab0
+      md5sum: dd87833db2f333dd2f9193c84edf23e5
     - path: results_test/RNAseq_report.html
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index ed0ffe17..a53cd1fd 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -70,13 +70,7 @@
     - path: results_test/differential_gene_expression/plots/PCA_plot.png
     - path: results_test/differential_gene_expression/plots/PCA_plot.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 8c6ed54d7faf0beb6d41afa9019c8857
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv
-      md5sum: 23ac7b3845e539fa5748fe58a920fd89
+      md5sum: 710bb4810bcf109e03b559cae54d5031
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 1b4ae63b60287794f8eb4d1fcaf04fa3
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv
-      md5sum: eb7e714867d0afb0f851b09ebf2c5a88
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png
+      md5sum: 68b329da9893e34099c7d8ad5cb9c940
     - path: results_test/RNAseq_report.html

From 9fea8e160b9d2388045423b91c4717f5a24317c0 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 10:42:01 +0100
Subject: [PATCH 029/124] switched venn test from full to small test

---
 tests/test.yml      | 6 ++++++
 tests/test_full.yml | 3 ---
 2 files changed, 6 insertions(+), 3 deletions(-)

diff --git a/tests/test.yml b/tests/test.yml
index 64ca823b..7ed85404 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -60,6 +60,9 @@
     - path: results_test/differential_gene_expression/plots/PCA_plot.pdf
     - path: results_test/differential_gene_expression/plots/PCA_plot.png
     - path: results_test/differential_gene_expression/plots/PCA_plot.svg
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png
+    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
@@ -82,6 +85,9 @@
       md5sum: 9318d57b86ba653f6183e61840a7943b
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
       md5sum: d5632f9240116f42de59f05b7e822a29
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
diff --git a/tests/test_full.yml b/tests/test_full.yml
index 02cd336a..d739d687 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -68,9 +68,6 @@
       md5sum: 7ee7cd938140217a863774c3689ff1af
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
       md5sum: 18873d39d839477337312cce0ec63398
-    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf
-    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png
-    - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf

From fe7095d1686267c5647b620d7dd2fe4365d6de41 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 13:57:32 +0100
Subject: [PATCH 030/124] renamed params, updated docu, added option of adding
 custom bg list

---
 assets/RNAseq_report.Rmd   | 25 +++++++++++++++++++------
 bin/Execute_report.R       |  6 ++++--
 docs/output.md             |  3 +++
 docs/usage.md              | 11 +++++++++++
 modules/local/report.nf    |  7 +++++--
 nextflow.config            |  3 ++-
 nextflow_schema.json       |  6 +++++-
 tests/test_batcheffect.yml |  2 +-
 workflows/rnadeseq.nf      |  4 +++-
 9 files changed, 53 insertions(+), 14 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 0cf78ce8..729a0af6 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -40,7 +40,8 @@ params:
     keytype: ''
     min_DEG_pathway: ''
     custom_gmt: ''
-    background_list: ''
+    set_background: ''
+    custom_background: ''
 
     #Additional args for the report
     path_proj_summary: ''
@@ -1554,6 +1555,16 @@ datasources <- c("KEGG", "REAC")
 # Default to 1 for the nb of genes DE in a pathway
 min_DEG_pathway <- as.integer(params$min_DEG_pathway)
 
+# If background list is to be used, load custom background list
+# if provided, else use notAllZero
+if (isProvided(params$set_background)) {
+    if (isProvided(params$custom_background)) {
+        custom_background <- readLines(params$custom_background)
+    } else {
+        custom_background <- rownames(counts(cds))[notAllZero]
+    }
+}
+
 # Contrast files
 contrast_files <- list.files('differential_gene_expression/DE_genes_tables/')
 path_contrasts <- 'differential_gene_expression/DE_genes_tables/'
@@ -1644,7 +1655,7 @@ for (file in contrast_files){
     q = as.character(DE_genes$Ensembl_ID)
 
     # gost query
-    if (params$background_list) {
+    if (isProvided(params$set_background)) {
         gostres <- gost(query=q,
             organism=gost_id,
             significant=TRUE,
@@ -1652,7 +1663,7 @@ for (file in contrast_files){
             sources=datasources,
             evcodes=TRUE,
             user_threshold=params$pval_threshold,
-            custom_bg=rownames(counts(cds))[notAllZero],
+            custom_bg=custom_background,
             domain_scope="custom_annotated"
         )
         
@@ -1716,7 +1727,7 @@ for (file in contrast_files){
             height=10, width=15, units="cm", dpi=300, limitsize=F)
 
         # If background list was used for main PA analysis, plot Venn diagram of the results and those from the additional PA without background list
-        if (params$background_list) {
+        if (isProvided(params$set_background)) {
             pathway_gostres_nobg <- gostres_nobg$result
             pathway_gostres_nobg <- as.data.frame(pathway_gostres_nobg[which(pathway_gostres_nobg$significant==TRUE),])
             pathway_gostres_nobg <- pathway_gostres_nobg[which(pathway_gostres_nobg$intersection_size>=min_DEG_pathway),]
@@ -1913,6 +1924,8 @@ After unzipping the results ZIP file, the pathway analysis results are stored in
 Inside the pathway analysis results folder, a subfolder for each contrast used for the differential gene expression analysis is found. Inside each contrast folder, there is the following output:
 <br><br><details>
 <summary>Expand/collapse</summary>
+- `*_gost_pathway_venn_diagram.pdf/png`
+  - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list.
 - `*_KEGG_pathway_enrichment_plot.pdf/png`
     - Barplots showing the proportion of differentially expressed genes in the pathway.
 - `KEGG_pathways/`
@@ -1957,14 +1970,14 @@ if (length(q_list) > 0) {
 
     #gost query
     
-    if (params$background_list) {
+    if (isProvided(params$set_background)) {
         gostres <- gost(query=q_list,
                     organism=gost_id,
                     significant=T,
                     correction_method="fdr",
                     sources=datasources,
                     user_threshold=params$pval_threshold,
-                    custom_bg=rownames(counts(cds))[notAllZero],
+                    custom_bg=custom_background,
                     domain_scope="custom_annotated"
         )
     } else {
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index 4246cea0..c14dba59 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -31,7 +31,8 @@ option_list = list(
     make_option(c("-i", "--species_library"), type="character", default=NULL, help="Library name. Example format: org.At.tair.db", metavar="character"),
     make_option(c("-u", "--keytype"), type="character", default=NULL, help="Keytype. Example format: TAIR (varies greatly depending on library!)", metavar="character"),
     make_option("--custom_gmt", type="character", default=NULL, help="Path to custom GMT file to query during pathway analysis.", metavar="character"),
-    make_option("--background_list", action="store_true", default=TRUE, help="Whether to use a background list for pathway analysis; if true, will only consider expressed genes (i.e. mean counts > 0) for PA."),
+    make_option("--set_background", action="store_true", default=TRUE, help="Whether to use a background list for pathway analysis; if true, will only consider expressed genes (i.e. mean counts > 0) for PA."),
+    make_option("--custom_background", type="character", default=NULL, help="Path to a custom background list TXT for pathway analysis; if provided, will only consider genes in that list for PA."),
     make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"),
 
     make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"),
@@ -77,7 +78,8 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 organism = opt$organism,
                                 species_library = opt$species_library,
                                 custom_gmt = opt$custom_gmt,
-                                background_list = opt$background_list,
+                                set_background = opt$set_background,
+                                custom_background = opt$custom_background,
                                 keytype = opt$keytype,
                                 min_DEG_pathway = opt$min_DEG_pathway,
 
diff --git a/docs/output.md b/docs/output.md
index ff953f84..9a5da113 100644
--- a/docs/output.md
+++ b/docs/output.md
@@ -49,6 +49,9 @@ Pathway analysis with [gProfileR](https://biit.cs.ut.ee/gprofiler/gost) R packag
 
 This directory contains the zipped pathway analysis results (`gProfileR.zip`). When unzipping them, a subfolder for each contrast used for the differential gene expression analysis is found. Inside each contrast folder, there is the following output:
 
+
+- `*_gost_pathway_venn_diagram.pdf/png`
+  - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list.
 - `*_KEGG_pathway_enrichment_plot.pdf/png`
   - Barplots showing the proportion of differentially expressed genes in the pathway.
 - `KEGG_pathways/`
diff --git a/docs/usage.md b/docs/usage.md
index 9cdcf800..81c4d2c6 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -45,6 +45,9 @@
   - [`--organism`](#--organism)
   - [`--species_library`](#--species_library)
   - [`--keytype`](#--keytype)
+  - [`--custom_gmt`](#--custom_gmt)
+  - [`--set_background`](#--set_background)
+  - [`--custom_background`](#--custom_background)  
   - [`--igenomes_base`](#--igenomes_base)
   - [`--igenomes_ignore`](#--igenomes_ignore)
 - [Job resources](#job-resources)
@@ -387,6 +390,14 @@ Which keytype to use for pathway analysis, e.g. ENSEMBL, not necessary if `--run
 
 Path to custom GMT file to use instead of querying against the live gprofiler database, not necessary if `--run_pathway_analysis = false`.
 
+### `--set_background`
+
+Whether to restrict pathway analysis to a background gene list (default: true, will restrict to those genes with an expression > 0 in at least one sample), not necessary if `--run_pathway_analysis = false`.
+
+### `--custom_background`
+
+Path to custom background TXT file with one gene ID per line to use as background genes, not necessary if `--run_pathway_analysis = false` or `--set_background = false`.
+
 ## Job resources
 
 ### Automatic resubmission
diff --git a/modules/local/report.nf b/modules/local/report.nf
index e59e45ea..87a853fa 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -18,6 +18,7 @@ process REPORT {
     path software_versions
     path multiqc
     path custom_gmt
+    path custom_background
 
     output:
     path "*.zip"
@@ -36,7 +37,8 @@ process REPORT {
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
-    def background_list_opt = params.background_list ? "--background_list TRUE" : "--background_list FALSE"
+    def set_background_opt = params.set_background ? "--set_background TRUE" : "--set_background FALSE"
+    def custom_background_opt = custom_background.name != 'NO_FILE7' ? "--custom_background $custom_background" : ''
 
     def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : ''
     def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : ''
@@ -70,7 +72,8 @@ process REPORT {
         $round_DE_opt \
         $pathway_opt \
         $custom_gmt_opt \
-        $background_list_opt \
+        $set_background_opt \
+        $custom_background_opt \
         --organism $params.organism \
         --species_library $params.species_library \
         --keytype $params.keytype \
diff --git a/nextflow.config b/nextflow.config
index 476d21cb..0452926f 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -37,7 +37,8 @@ params {
     run_pathway_analysis = false
     min_DEG_pathway = 1
     custom_gmt = 'NO_FILE3'
-    background_list = true
+    set_background = true
+    custom_background = 'NO_FILE7'
 
     // Additional args for the report
     project_summary = null
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 2f5ae149..0f7a75cc 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -124,11 +124,15 @@
                     "type": "string",
                     "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true."
                 },
-                "background_list": {
+                "set_background": {
                     "type": "boolean",
                     "default": true,
                     "description": "Whether to limit pathway analysis to a background list of genes (i.e. those genes expressed in at least one sample), default = true."
                 },
+                "custom_background": {
+                    "type": "string",
+                    "description": "Path to custom background TXT file for gost query containing a gene ID in each line, not necessary if --skip_pathway_analysis = true or --set_background = false."
+                },
                 "min_DEG_pathway": {
                     "type": "integer",
                     "default": 1,
diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml
index 7afa1153..216f3301 100644
--- a/tests/test_batcheffect.yml
+++ b/tests/test_batcheffect.yml
@@ -1,5 +1,5 @@
 - name: Run test profile with batcheffect correction
-  command: nextflow run main.nf -profile test_batcheffect,docker --citest true --round_DE 3 --background_list false
+  command: nextflow run main.nf -profile test_batcheffect,docker --citest true --round_DE 3 --set_background false
   tags:
     - test_batcheffect
   files:
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
index 07c634ff..58bb8cd0 100644
--- a/workflows/rnadeseq.nf
+++ b/workflows/rnadeseq.nf
@@ -34,6 +34,7 @@ ch_relevel = Channel.fromPath(params.relevel)
 ch_genes = Channel.fromPath(params.genelist)
 ch_multiqc_file = Channel.fromPath(params.multiqc)
 ch_custom_gmt = Channel.fromPath(params.custom_gmt)
+ch_custom_background = Channel.fromPath(params.custom_background)
 ch_proj_summary_file = Channel.fromPath(params.project_summary)
 ch_softwareversions_file = Channel.fromPath(params.software_versions)
 
@@ -98,7 +99,8 @@ workflow RNADESEQ {
         ch_proj_summary_file,
         ch_softwareversions_file,
         ch_multiqc_file,
-        ch_custom_gmt
+        ch_custom_gmt,
+        ch_custom_background
     )
 
     //TODO: Enable this:

From 0e8c9e3b903e1fbd742f49be75e4d78bc90bac72 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 15 Mar 2023 14:55:59 +0100
Subject: [PATCH 031/124] prettier

---
 docs/output.md | 1 -
 docs/usage.md  | 2 +-
 2 files changed, 1 insertion(+), 2 deletions(-)

diff --git a/docs/output.md b/docs/output.md
index 9a5da113..bdc278d0 100644
--- a/docs/output.md
+++ b/docs/output.md
@@ -49,7 +49,6 @@ Pathway analysis with [gProfileR](https://biit.cs.ut.ee/gprofiler/gost) R packag
 
 This directory contains the zipped pathway analysis results (`gProfileR.zip`). When unzipping them, a subfolder for each contrast used for the differential gene expression analysis is found. Inside each contrast folder, there is the following output:
 
-
 - `*_gost_pathway_venn_diagram.pdf/png`
   - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list.
 - `*_KEGG_pathway_enrichment_plot.pdf/png`
diff --git a/docs/usage.md b/docs/usage.md
index 81c4d2c6..93002a3e 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -47,7 +47,7 @@
   - [`--keytype`](#--keytype)
   - [`--custom_gmt`](#--custom_gmt)
   - [`--set_background`](#--set_background)
-  - [`--custom_background`](#--custom_background)  
+  - [`--custom_background`](#--custom_background)
   - [`--igenomes_base`](#--igenomes_base)
   - [`--igenomes_ignore`](#--igenomes_ignore)
 - [Job resources](#job-resources)

From ceaaf5bfeb4a7b48aee06127b2bcf31a8c903019 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 17 Mar 2023 09:27:07 +0100
Subject: [PATCH 032/124] updated dockerfile and git ci for the ghcr changes

---
 .github/workflows/ci.yml | 12 ++++++------
 Dockerfile               |  7 ++++---
 2 files changed, 10 insertions(+), 9 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index f6ee4de7..3a29fb20 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -36,13 +36,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
-          docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
+          docker pull ghcr.io/qbic-pipelines/rnadeseq:dev
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
@@ -88,13 +88,13 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:dev
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev
 
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
-          docker pull qbicpipelines/rnadeseq:dev
-          docker tag qbicpipelines/rnadeseq:dev qbicpipelines/rnadeseq:dev
+          docker pull ghcr.io/qbic-pipelines/rnadeseq:dev
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
diff --git a/Dockerfile b/Dockerfile
index aef20cc2..37680a0c 100644
--- a/Dockerfile
+++ b/Dockerfile
@@ -1,6 +1,8 @@
 FROM condaforge/mambaforge
-LABEL authors="Gisela Gabernet, Alexander Peltzer" \
-    description="Docker image containing all requirements for qbic-pipelines/rnadeseq pipeline"
+LABEL org.opencontainers.image.source=https://github.com/qbic-pipelines/rnadeseq
+LABEL org.opencontainers.image.description="Docker image containing all requirements for qbic-pipelines/rnadeseq pipeline"
+LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer"
+LABEL org.opencontainers.image.licenses=MIT
 COPY environment.yml /
 #RUN conda install -c conda-forge mamba
 RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \
@@ -14,4 +16,3 @@ RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadese
 # Instruct R processes to use these empty files instead of clashing with a local config
 RUN touch .Rprofile
 RUN touch .Renviron
-

From 69e9fce49ff1d040f4389a873885da3dbd301335 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 17 Mar 2023 10:05:56 +0100
Subject: [PATCH 033/124] updated push_dockerhub

---
 .github/workflows/push_dockerhub.yml | 24 +++++++++++-------------
 1 file changed, 11 insertions(+), 13 deletions(-)

diff --git a/.github/workflows/push_dockerhub.yml b/.github/workflows/push_dockerhub.yml
index e15120d9..da1b5b07 100644
--- a/.github/workflows/push_dockerhub.yml
+++ b/.github/workflows/push_dockerhub.yml
@@ -14,27 +14,25 @@ jobs:
     runs-on: ubuntu-latest
     # Only run for the qbicpipelines repo, for releases and merged PRs
     if: ${{ github.repository == 'qbic-pipelines/rnadeseq' }}
-    env:
-      DOCKERHUB_USERNAME: ${{ secrets.DOCKERHUB_USERNAME }}
-      DOCKERHUB_PASS: ${{ secrets.DOCKERHUB_PASS }}
     steps:
       - name: Check out pipeline code
         uses: actions/checkout@v2
 
       - name: Build new docker image
-        run: docker build --no-cache . -t qbicpipelines/rnadeseq:latest
+        run: docker build --no-cache . -t ghcr.io/qbicpipelines/rnadeseq:latest
 
-      - name: Push Docker image to DockerHub (dev)
+      - name: Log in to registry
+        # Update the personal access token to GITHUB_TOKEN
+        run: echo "${{ secrets.GITHUB_TOKEN }}" | docker login ghcr.io -u $ --password-stdin
+
+      - name: Push image to GHCR (dev)
         if: ${{ github.event_name == 'push' }}
         run: |
-          echo "$DOCKERHUB_PASS" | docker login --username "$DOCKERHUB_USERNAME" --password-stdin
-          docker tag qbicpipelines/rnadeseq:latest qbicpipelines/rnadeseq:dev
-          docker push qbicpipelines/rnadeseq:dev
+          docker tag ghcr.io/qbicpipelines/rnadeseq:latest ghcr.io/qbicpipelines/rnadeseq:dev
+          docker push ghcr.io/qbicpipelines/rnadeseq:dev
 
-      - name: Push Docker image to DockerHub (release)
+      - name: Push image to GHCR (release)
         if: ${{ github.event_name == 'release' }}
         run: |
-          echo "$DOCKERHUB_PASS" | docker login -u "$DOCKERHUB_USERNAME" --password-stdin
-          docker push qbicpipelines/rnadeseq:latest
-          docker tag qbicpipelines/rnadeseq:latest qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}
-          docker push qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}
+          docker tag ghcr.io/qbicpipelines/rnadeseq:latest ghcr.io/qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}
+          docker push ghcr.io/qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}

From 806517c36db0344a3b0c873b5504eb9d0af1e4f9 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 17 Mar 2023 11:24:21 +0100
Subject: [PATCH 034/124] updated changelog

---
 CHANGELOG.md | 1 +
 1 file changed, 1 insertion(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 26b67dea..57724697 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -13,6 +13,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#203](https://github.com/qbic-pipelines/rnadeseq/pull/203) Switched from Dockerhub to GHCR
 - [#200](https://github.com/qbic-pipelines/rnadeseq/pull/200) Made software_versions optional
 - [#198](https://github.com/qbic-pipelines/rnadeseq/pull/198) Changed heatmaps to scale in size automatically
 

From 43bcde96f2883a96c653c09724f2d9efe05429e8 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Thu, 23 Mar 2023 10:14:44 +0100
Subject: [PATCH 035/124] Update Dockerfile

Co-authored-by: Gisela Gabernet <gisela.gabernet@gmail.com>
---
 Dockerfile | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/Dockerfile b/Dockerfile
index 37680a0c..4f8ce7fe 100644
--- a/Dockerfile
+++ b/Dockerfile
@@ -1,7 +1,7 @@
 FROM condaforge/mambaforge
 LABEL org.opencontainers.image.source=https://github.com/qbic-pipelines/rnadeseq
 LABEL org.opencontainers.image.description="Docker image containing all requirements for qbic-pipelines/rnadeseq pipeline"
-LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer"
+LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oskar Wacker"
 LABEL org.opencontainers.image.licenses=MIT
 COPY environment.yml /
 #RUN conda install -c conda-forge mamba

From f9f6c4be72fae20aa7ecc213104e699d7f521184 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 23 Mar 2023 10:24:45 +0100
Subject: [PATCH 036/124] renamed push_dockerhub.yml to
 push_github_container_reg.yml; changed container in module

---
 .../{push_dockerhub.yml => push_github_container_reg.yml}       | 2 +-
 modules/local/report.nf                                         | 2 +-
 2 files changed, 2 insertions(+), 2 deletions(-)
 rename .github/workflows/{push_dockerhub.yml => push_github_container_reg.yml} (95%)

diff --git a/.github/workflows/push_dockerhub.yml b/.github/workflows/push_github_container_reg.yml
similarity index 95%
rename from .github/workflows/push_dockerhub.yml
rename to .github/workflows/push_github_container_reg.yml
index da1b5b07..c4e81bac 100644
--- a/.github/workflows/push_dockerhub.yml
+++ b/.github/workflows/push_github_container_reg.yml
@@ -1,5 +1,5 @@
 name: Docker push
-# This builds the docker image and pushes it to DockerHub
+# This builds the docker image and pushes it to GHCR
 # Runs on qbic-pipelines repo releases and push event to 'dev' branch (PR merges)
 on:
   push:
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 87a853fa..1e6d8dc5 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -1,6 +1,6 @@
 process REPORT {
 
-    container 'qbicpipelines/rnadeseq:dev'
+    container 'ghcr.io/qbic-pipelines/rnadeseq:dev'
 
     input:
     path gene_counts

From 5fb1b58bc098cbf8eb706095d5026dceca8b055f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 3 Apr 2023 08:55:21 +0200
Subject: [PATCH 037/124] Fixed container name

---
 .github/workflows/push_github_container_reg.yml | 12 ++++++------
 1 file changed, 6 insertions(+), 6 deletions(-)

diff --git a/.github/workflows/push_github_container_reg.yml b/.github/workflows/push_github_container_reg.yml
index c4e81bac..b8934ae3 100644
--- a/.github/workflows/push_github_container_reg.yml
+++ b/.github/workflows/push_github_container_reg.yml
@@ -12,14 +12,14 @@ jobs:
   push_dockerhub:
     name: Push new Docker image to Docker Hub
     runs-on: ubuntu-latest
-    # Only run for the qbicpipelines repo, for releases and merged PRs
+    # Only run for the qbic-pipelines repo, for releases and merged PRs
     if: ${{ github.repository == 'qbic-pipelines/rnadeseq' }}
     steps:
       - name: Check out pipeline code
         uses: actions/checkout@v2
 
       - name: Build new docker image
-        run: docker build --no-cache . -t ghcr.io/qbicpipelines/rnadeseq:latest
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:latest
 
       - name: Log in to registry
         # Update the personal access token to GITHUB_TOKEN
@@ -28,11 +28,11 @@ jobs:
       - name: Push image to GHCR (dev)
         if: ${{ github.event_name == 'push' }}
         run: |
-          docker tag ghcr.io/qbicpipelines/rnadeseq:latest ghcr.io/qbicpipelines/rnadeseq:dev
-          docker push ghcr.io/qbicpipelines/rnadeseq:dev
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:latest ghcr.io/qbic-pipelines/rnadeseq:dev
+          docker push ghcr.io/qbic-pipelines/rnadeseq:dev
 
       - name: Push image to GHCR (release)
         if: ${{ github.event_name == 'release' }}
         run: |
-          docker tag ghcr.io/qbicpipelines/rnadeseq:latest ghcr.io/qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}
-          docker push ghcr.io/qbicpipelines/rnadeseq:${{ github.event.release.tag_name }}
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:latest ghcr.io/qbic-pipelines/rnadeseq:${{ github.event.release.tag_name }}
+          docker push ghcr.io/qbic-pipelines/rnadeseq:${{ github.event.release.tag_name }}

From 849eb960c6e6abfd536530df06efea54a0c28046 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 19 Apr 2023 10:04:28 +0200
Subject: [PATCH 038/124] Changed relevel github path to qbic-pipelines

---
 CHANGELOG.md             | 1 +
 conf/test_relevel.config | 2 +-
 2 files changed, 2 insertions(+), 1 deletion(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 57724697..caf0c341 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -13,6 +13,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#204](https://github.com/qbic-pipelines/rnadeseq/pull/204) Changed relevel path in test_relevel.config to the qbic-pipelines repo
 - [#203](https://github.com/qbic-pipelines/rnadeseq/pull/203) Switched from Dockerhub to GHCR
 - [#200](https://github.com/qbic-pipelines/rnadeseq/pull/200) Made software_versions optional
 - [#198](https://github.com/qbic-pipelines/rnadeseq/pull/198) Changed heatmaps to scale in size automatically
diff --git a/conf/test_relevel.config b/conf/test_relevel.config
index e43c6dc3..4a07c212 100644
--- a/conf/test_relevel.config
+++ b/conf/test_relevel.config
@@ -18,7 +18,7 @@ params {
     input = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/Sample_preparations.tsv'
     gene_counts = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/merged_gene_counts.txt'
     model = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/design.txt'
-    relevel = 'https://raw.githubusercontent.com/SusiJo/rnadeseq/dev/testdata/relevel.tsv'
+    relevel = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/relevel.tsv'
     genelist = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/requested_genes.txt'
     //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml'
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'

From 375172597ba847596d0c2cee46e48c121050d270 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 07:37:18 +0200
Subject: [PATCH 039/124] Initial template commit

---
 .devcontainer/devcontainer.json               |  27 +
 .editorconfig                                 |  24 +
 .gitattributes                                |   4 +
 .github/.dockstore.yml                        |   6 +
 .github/CONTRIBUTING.md                       | 119 ++++
 .github/ISSUE_TEMPLATE/bug_report.yml         |  50 ++
 .github/ISSUE_TEMPLATE/config.yml             |   7 +
 .github/ISSUE_TEMPLATE/feature_request.yml    |  11 +
 .github/PULL_REQUEST_TEMPLATE.md              |  24 +
 .github/workflows/awsfulltest.yml             |  34 ++
 .github/workflows/awstest.yml                 |  29 +
 .github/workflows/branch.yml                  |  44 ++
 .github/workflows/ci.yml                      |  43 ++
 .github/workflows/fix-linting.yml             |  55 ++
 .github/workflows/linting.yml                 | 108 ++++
 .github/workflows/linting_comment.yml         |  28 +
 .gitignore                                    |   8 +
 .gitpod.yml                                   |  14 +
 .nf-core.yml                                  |   1 +
 .prettierignore                               |  12 +
 .prettierrc.yml                               |   1 +
 CHANGELOG.md                                  |  16 +
 CITATIONS.md                                  |  35 ++
 CODE_OF_CONDUCT.md                            | 111 ++++
 LICENSE                                       |  21 +
 README.md                                     |  92 +++
 assets/adaptivecard.json                      |  67 +++
 assets/email_template.html                    |  53 ++
 assets/email_template.txt                     |  39 ++
 assets/methods_description_template.yml       |  25 +
 assets/multiqc_config.yml                     |  13 +
 assets/nf-core-rnadeseq_logo_light.png        | Bin 0 -> 11674 bytes
 assets/samplesheet.csv                        |   3 +
 assets/schema_input.json                      |  36 ++
 assets/sendmail_template.txt                  |  53 ++
 assets/slackreport.json                       |  34 ++
 bin/check_samplesheet.py                      | 262 +++++++++
 conf/base.config                              |  65 +++
 conf/igenomes.config                          | 432 ++++++++++++++
 conf/modules.config                           |  41 ++
 conf/test.config                              |  29 +
 conf/test_full.config                         |  24 +
 docs/README.md                                |  10 +
 docs/images/mqc_fastqc_adapter.png            | Bin 0 -> 23458 bytes
 docs/images/mqc_fastqc_counts.png             | Bin 0 -> 33918 bytes
 docs/images/mqc_fastqc_quality.png            | Bin 0 -> 55769 bytes
 docs/images/nf-core-rnadeseq_logo_dark.png    | Bin 0 -> 75896 bytes
 docs/images/nf-core-rnadeseq_logo_light.png   | Bin 0 -> 75779 bytes
 docs/output.md                                |  68 +++
 docs/usage.md                                 | 270 +++++++++
 lib/NfcoreSchema.groovy                       | 528 ++++++++++++++++++
 lib/NfcoreTemplate.groovy                     | 336 +++++++++++
 lib/Utils.groovy                              |  47 ++
 lib/WorkflowMain.groovy                       |  99 ++++
 lib/WorkflowRnadeseq.groovy                   |  77 +++
 lib/nfcore_external_java_deps.jar             | Bin 0 -> 2291171 bytes
 main.nf                                       |  64 +++
 modules.json                                  |  27 +
 modules/local/samplesheet_check.nf            |  31 +
 .../custom/dumpsoftwareversions/main.nf       |  24 +
 .../custom/dumpsoftwareversions/meta.yml      |  34 ++
 .../templates/dumpsoftwareversions.py         | 102 ++++
 modules/nf-core/fastqc/main.nf                |  51 ++
 modules/nf-core/fastqc/meta.yml               |  52 ++
 modules/nf-core/multiqc/main.nf               |  53 ++
 modules/nf-core/multiqc/meta.yml              |  55 ++
 nextflow.config                               | 234 ++++++++
 nextflow_schema.json                          | 287 ++++++++++
 pyproject.toml                                |  10 +
 subworkflows/local/input_check.nf             |  44 ++
 workflows/rnadeseq.nf                         | 132 +++++
 71 files changed, 4735 insertions(+)
 create mode 100644 .devcontainer/devcontainer.json
 create mode 100644 .editorconfig
 create mode 100644 .gitattributes
 create mode 100644 .github/.dockstore.yml
 create mode 100644 .github/CONTRIBUTING.md
 create mode 100644 .github/ISSUE_TEMPLATE/bug_report.yml
 create mode 100644 .github/ISSUE_TEMPLATE/config.yml
 create mode 100644 .github/ISSUE_TEMPLATE/feature_request.yml
 create mode 100644 .github/PULL_REQUEST_TEMPLATE.md
 create mode 100644 .github/workflows/awsfulltest.yml
 create mode 100644 .github/workflows/awstest.yml
 create mode 100644 .github/workflows/branch.yml
 create mode 100644 .github/workflows/ci.yml
 create mode 100644 .github/workflows/fix-linting.yml
 create mode 100644 .github/workflows/linting.yml
 create mode 100644 .github/workflows/linting_comment.yml
 create mode 100644 .gitignore
 create mode 100644 .gitpod.yml
 create mode 100644 .nf-core.yml
 create mode 100644 .prettierignore
 create mode 100644 .prettierrc.yml
 create mode 100644 CHANGELOG.md
 create mode 100644 CITATIONS.md
 create mode 100644 CODE_OF_CONDUCT.md
 create mode 100644 LICENSE
 create mode 100644 README.md
 create mode 100644 assets/adaptivecard.json
 create mode 100644 assets/email_template.html
 create mode 100644 assets/email_template.txt
 create mode 100644 assets/methods_description_template.yml
 create mode 100644 assets/multiqc_config.yml
 create mode 100644 assets/nf-core-rnadeseq_logo_light.png
 create mode 100644 assets/samplesheet.csv
 create mode 100644 assets/schema_input.json
 create mode 100644 assets/sendmail_template.txt
 create mode 100644 assets/slackreport.json
 create mode 100755 bin/check_samplesheet.py
 create mode 100644 conf/base.config
 create mode 100644 conf/igenomes.config
 create mode 100644 conf/modules.config
 create mode 100644 conf/test.config
 create mode 100644 conf/test_full.config
 create mode 100644 docs/README.md
 create mode 100755 docs/images/mqc_fastqc_adapter.png
 create mode 100755 docs/images/mqc_fastqc_counts.png
 create mode 100755 docs/images/mqc_fastqc_quality.png
 create mode 100644 docs/images/nf-core-rnadeseq_logo_dark.png
 create mode 100644 docs/images/nf-core-rnadeseq_logo_light.png
 create mode 100644 docs/output.md
 create mode 100644 docs/usage.md
 create mode 100755 lib/NfcoreSchema.groovy
 create mode 100755 lib/NfcoreTemplate.groovy
 create mode 100644 lib/Utils.groovy
 create mode 100755 lib/WorkflowMain.groovy
 create mode 100755 lib/WorkflowRnadeseq.groovy
 create mode 100644 lib/nfcore_external_java_deps.jar
 create mode 100644 main.nf
 create mode 100644 modules.json
 create mode 100644 modules/local/samplesheet_check.nf
 create mode 100644 modules/nf-core/custom/dumpsoftwareversions/main.nf
 create mode 100644 modules/nf-core/custom/dumpsoftwareversions/meta.yml
 create mode 100755 modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
 create mode 100644 modules/nf-core/fastqc/main.nf
 create mode 100644 modules/nf-core/fastqc/meta.yml
 create mode 100644 modules/nf-core/multiqc/main.nf
 create mode 100644 modules/nf-core/multiqc/meta.yml
 create mode 100644 nextflow.config
 create mode 100644 nextflow_schema.json
 create mode 100644 pyproject.toml
 create mode 100644 subworkflows/local/input_check.nf
 create mode 100644 workflows/rnadeseq.nf

diff --git a/.devcontainer/devcontainer.json b/.devcontainer/devcontainer.json
new file mode 100644
index 00000000..ea27a584
--- /dev/null
+++ b/.devcontainer/devcontainer.json
@@ -0,0 +1,27 @@
+{
+    "name": "nfcore",
+    "image": "nfcore/gitpod:latest",
+    "remoteUser": "gitpod",
+
+    // Configure tool-specific properties.
+    "customizations": {
+        // Configure properties specific to VS Code.
+        "vscode": {
+            // Set *default* container specific settings.json values on container create.
+            "settings": {
+                "python.defaultInterpreterPath": "/opt/conda/bin/python",
+                "python.linting.enabled": true,
+                "python.linting.pylintEnabled": true,
+                "python.formatting.autopep8Path": "/opt/conda/bin/autopep8",
+                "python.formatting.yapfPath": "/opt/conda/bin/yapf",
+                "python.linting.flake8Path": "/opt/conda/bin/flake8",
+                "python.linting.pycodestylePath": "/opt/conda/bin/pycodestyle",
+                "python.linting.pydocstylePath": "/opt/conda/bin/pydocstyle",
+                "python.linting.pylintPath": "/opt/conda/bin/pylint"
+            },
+
+            // Add the IDs of extensions you want installed when the container is created.
+            "extensions": ["ms-python.python", "ms-python.vscode-pylance", "nf-core.nf-core-extensionpack"]
+        }
+    }
+}
diff --git a/.editorconfig b/.editorconfig
new file mode 100644
index 00000000..b78de6e6
--- /dev/null
+++ b/.editorconfig
@@ -0,0 +1,24 @@
+root = true
+
+[*]
+charset = utf-8
+end_of_line = lf
+insert_final_newline = true
+trim_trailing_whitespace = true
+indent_size = 4
+indent_style = space
+
+[*.{md,yml,yaml,html,css,scss,js,cff}]
+indent_size = 2
+
+# These files are edited and tested upstream in nf-core/modules
+[/modules/nf-core/**]
+charset = unset
+end_of_line = unset
+insert_final_newline = unset
+trim_trailing_whitespace = unset
+indent_style = unset
+indent_size = unset
+
+[/assets/email*]
+indent_size = unset
diff --git a/.gitattributes b/.gitattributes
new file mode 100644
index 00000000..7a2dabc2
--- /dev/null
+++ b/.gitattributes
@@ -0,0 +1,4 @@
+*.config linguist-language=nextflow
+*.nf.test linguist-language=nextflow
+modules/nf-core/** linguist-generated
+subworkflows/nf-core/** linguist-generated
diff --git a/.github/.dockstore.yml b/.github/.dockstore.yml
new file mode 100644
index 00000000..191fabd2
--- /dev/null
+++ b/.github/.dockstore.yml
@@ -0,0 +1,6 @@
+# Dockstore config version, not pipeline version
+version: 1.2
+workflows:
+  - subclass: nfl
+    primaryDescriptorPath: /nextflow.config
+    publish: True
diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md
new file mode 100644
index 00000000..dc88c9b5
--- /dev/null
+++ b/.github/CONTRIBUTING.md
@@ -0,0 +1,119 @@
+# nf-core/rnadeseq: Contributing Guidelines
+
+Hi there!
+Many thanks for taking an interest in improving nf-core/rnadeseq.
+
+We try to manage the required tasks for nf-core/rnadeseq using GitHub issues, you probably came to this page when creating one.
+Please use the pre-filled template to save time.
+
+However, don't be put off by this template - other more general issues and suggestions are welcome!
+Contributions to the code are even more welcome ;)
+
+> If you need help using or modifying nf-core/rnadeseq then the best place to ask is on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)).
+
+## Contribution workflow
+
+If you'd like to write some code for nf-core/rnadeseq, the standard workflow is as follows:
+
+1. Check that there isn't already an issue about your idea in the [nf-core/rnadeseq issues](https://github.com/nf-core/rnadeseq/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this
+2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/rnadeseq repository](https://github.com/nf-core/rnadeseq) to your GitHub account
+3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions)
+4. Use `nf-core schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10).
+5. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged
+
+If you're not used to this workflow with git, you can start with some [docs from GitHub](https://help.github.com/en/github/collaborating-with-issues-and-pull-requests) or even their [excellent `git` resources](https://try.github.io/).
+
+## Tests
+
+When you create a pull request with changes, [GitHub Actions](https://github.com/features/actions) will run automatic tests.
+Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then.
+
+There are typically two types of tests that run:
+
+### Lint tests
+
+`nf-core` has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to.
+To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint <pipeline-directory>` command.
+
+If any failures or warnings are encountered, please follow the listed URL for more documentation.
+
+### Pipeline tests
+
+Each `nf-core` pipeline should be set up with a minimal set of test-data.
+`GitHub Actions` then runs the pipeline on this data to ensure that it exits successfully.
+If there are any failures then the automated tests fail.
+These tests are run both with the latest available version of `Nextflow` and also the minimum required version that is stated in the pipeline code.
+
+## Patch
+
+:warning: Only in the unlikely and regretful event of a release happening with a bug.
+
+- On your own fork, make a new branch `patch` based on `upstream/master`.
+- Fix the bug, and bump version (X.Y.Z+1).
+- A PR should be made on `master` from patch to directly this particular bug.
+
+## Getting help
+
+For further information/help, please consult the [nf-core/rnadeseq documentation](https://nf-co.re/rnadeseq/usage) and don't hesitate to get in touch on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)).
+
+## Pipeline contribution conventions
+
+To make the nf-core/rnadeseq code and processing logic more understandable for new contributors and to ensure quality, we semi-standardise the way the code and other contributions are written.
+
+### Adding a new step
+
+If you wish to contribute a new step, please use the following coding standards:
+
+1. Define the corresponding input channel into your new process from the expected previous process channel
+2. Write the process block (see below).
+3. Define the output channel if needed (see below).
+4. Add any new parameters to `nextflow.config` with a default (see below).
+5. Add any new parameters to `nextflow_schema.json` with help text (via the `nf-core schema build` tool).
+6. Add sanity checks and validation for all relevant parameters.
+7. Perform local tests to validate that the new code works as expected.
+8. If applicable, add a new test command in `.github/workflow/ci.yml`.
+9. Update MultiQC config `assets/multiqc_config.yml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module.
+10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`.
+
+### Default values
+
+Parameters should be initialised / defined with default values in `nextflow.config` under the `params` scope.
+
+Once there, use `nf-core schema build` to add to `nextflow_schema.json`.
+
+### Default processes resource requirements
+
+Sensible defaults for process resource requirements (CPUs / memory / time) for a process should be defined in `conf/base.config`. These should generally be specified generic with `withLabel:` selectors so they can be shared across multiple processes/steps of the pipeline. A nf-core standard set of labels that should be followed where possible can be seen in the [nf-core pipeline template](https://github.com/nf-core/tools/blob/master/nf_core/pipeline-template/conf/base.config), which has the default process as a single core-process, and then different levels of multi-core configurations for increasingly large memory requirements defined with standardised labels.
+
+The process resources can be passed on to the tool dynamically within the process with the `${task.cpu}` and `${task.memory}` variables in the `script:` block.
+
+### Naming schemes
+
+Please use the following naming schemes, to make it easy to understand what is going where.
+
+- initial process channel: `ch_output_from_<process>`
+- intermediate and terminal channels: `ch_<previousprocess>_for_<nextprocess>`
+
+### Nextflow version bumping
+
+If you are using a new feature from core Nextflow, you may bump the minimum required version of nextflow in the pipeline with: `nf-core bump-version --nextflow . [min-nf-version]`
+
+### Images and figures
+
+For overview images and other documents we follow the nf-core [style guidelines and examples](https://nf-co.re/developers/design_guidelines).
+
+## GitHub Codespaces
+
+This repo includes a devcontainer configuration which will create a GitHub Codespaces for Nextflow development! This is an online developer environment that runs in your browser, complete with VSCode and a terminal.
+
+To get started:
+
+- Open the repo in [Codespaces](https://github.com/nf-core/rnadeseq/codespaces)
+- Tools installed
+  - nf-core
+  - Nextflow
+
+Devcontainer specs:
+
+- [DevContainer config](.devcontainer/devcontainer.json)
+- [Dockerfile](.devcontainer/Dockerfile)
diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml
new file mode 100644
index 00000000..5fb0efb4
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/bug_report.yml
@@ -0,0 +1,50 @@
+name: Bug report
+description: Report something that is broken or incorrect
+labels: bug
+body:
+  - type: markdown
+    attributes:
+      value: |
+        Before you post this issue, please check the documentation:
+
+        - [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting)
+        - [nf-core/rnadeseq pipeline documentation](https://nf-co.re/rnadeseq/usage)
+
+  - type: textarea
+    id: description
+    attributes:
+      label: Description of the bug
+      description: A clear and concise description of what the bug is.
+    validations:
+      required: true
+
+  - type: textarea
+    id: command_used
+    attributes:
+      label: Command used and terminal output
+      description: Steps to reproduce the behaviour. Please paste the command you used to launch the pipeline and the output from your terminal.
+      render: console
+      placeholder: |
+        $ nextflow run ...
+
+        Some output where something broke
+
+  - type: textarea
+    id: files
+    attributes:
+      label: Relevant files
+      description: |
+        Please drag and drop the relevant files here. Create a `.zip` archive if the extension is not allowed.
+        Your verbose log file `.nextflow.log` is often useful _(this is a hidden file in the directory where you launched the pipeline)_ as well as custom Nextflow configuration files.
+
+  - type: textarea
+    id: system
+    attributes:
+      label: System information
+      description: |
+        * Nextflow version _(eg. 22.10.1)_
+        * Hardware _(eg. HPC, Desktop, Cloud)_
+        * Executor _(eg. slurm, local, awsbatch)_
+        * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter or Charliecloud)_
+        * OS _(eg. CentOS Linux, macOS, Linux Mint)_
+        * Version of nf-core/rnadeseq _(eg. 1.1, 1.5, 1.8.2)_
diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml
new file mode 100644
index 00000000..1c032cae
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/config.yml
@@ -0,0 +1,7 @@
+contact_links:
+  - name: Join nf-core
+    url: https://nf-co.re/join
+    about: Please join the nf-core community here
+  - name: "Slack #rnadeseq channel"
+    url: https://nfcore.slack.com/channels/rnadeseq
+    about: Discussion about the nf-core/rnadeseq pipeline
diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml
new file mode 100644
index 00000000..6bd51f6c
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/feature_request.yml
@@ -0,0 +1,11 @@
+name: Feature request
+description: Suggest an idea for the nf-core/rnadeseq pipeline
+labels: enhancement
+body:
+  - type: textarea
+    id: description
+    attributes:
+      label: Description of feature
+      description: Please describe your suggestion for a new feature. It might help to describe a problem or use case, plus any alternatives that you have considered.
+    validations:
+      required: true
diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md
new file mode 100644
index 00000000..35658cf3
--- /dev/null
+++ b/.github/PULL_REQUEST_TEMPLATE.md
@@ -0,0 +1,24 @@
+<!--
+# nf-core/rnadeseq pull request
+
+Many thanks for contributing to nf-core/rnadeseq!
+
+Please fill in the appropriate checklist below (delete whatever is not relevant).
+These are the most common things requested on pull requests (PRs).
+
+Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release.
+
+Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/rnadeseq/tree/master/.github/CONTRIBUTING.md)
+-->
+
+## PR checklist
+
+- [ ] This comment contains a description of changes (with reason).
+- [ ] If you've fixed a bug or added code that should be tested, add tests!
+- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/rnadeseq/tree/master/.github/CONTRIBUTING.md)- [ ] If necessary, also make a PR on the nf-core/rnadeseq _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository.
+- [ ] Make sure your code lints (`nf-core lint`).
+- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir <OUTDIR>`).
+- [ ] Usage Documentation in `docs/usage.md` is updated.
+- [ ] Output Documentation in `docs/output.md` is updated.
+- [ ] `CHANGELOG.md` is updated.
+- [ ] `README.md` is updated (including new tool citations and authors/contributors).
diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml
new file mode 100644
index 00000000..ca110b1a
--- /dev/null
+++ b/.github/workflows/awsfulltest.yml
@@ -0,0 +1,34 @@
+name: nf-core AWS full size tests
+# This workflow is triggered on published releases.
+# It can be additionally triggered manually with GitHub actions workflow dispatch button.
+# It runs the -profile 'test_full' on AWS batch
+
+on:
+  release:
+    types: [published]
+  workflow_dispatch:
+jobs:
+  run-tower:
+    name: Run AWS full tests
+    if: github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      - name: Launch workflow via tower
+        uses: nf-core/tower-action@v3
+        # TODO nf-core: You can customise AWS full pipeline tests as required
+        # Add full size test data (but still relatively small datasets for few samples)
+        # on the `test_full.config` test runs with only one set of parameters
+        with:
+          workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }}
+          access_token: ${{ secrets.TOWER_ACCESS_TOKEN }}
+          compute_env: ${{ secrets.TOWER_COMPUTE_ENV }}
+          workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }}
+          parameters: |
+            {
+              "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-${{ github.sha }}"
+            }
+          profiles: test_full,aws_tower
+      - uses: actions/upload-artifact@v3
+        with:
+          name: Tower debug log file
+          path: tower_action_*.log
diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml
new file mode 100644
index 00000000..859570f1
--- /dev/null
+++ b/.github/workflows/awstest.yml
@@ -0,0 +1,29 @@
+name: nf-core AWS test
+# This workflow can be triggered manually with the GitHub actions workflow dispatch button.
+# It runs the -profile 'test' on AWS batch
+
+on:
+  workflow_dispatch:
+jobs:
+  run-tower:
+    name: Run AWS tests
+    if: github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      # Launch workflow using Tower CLI tool action
+      - name: Launch workflow via tower
+        uses: nf-core/tower-action@v3
+        with:
+          workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }}
+          access_token: ${{ secrets.TOWER_ACCESS_TOKEN }}
+          compute_env: ${{ secrets.TOWER_COMPUTE_ENV }}
+          workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }}
+          parameters: |
+            {
+              "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-test-${{ github.sha }}"
+            }
+          profiles: test,aws_tower
+      - uses: actions/upload-artifact@v3
+        with:
+          name: Tower debug log file
+          path: tower_action_*.log
diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml
new file mode 100644
index 00000000..2798fd8c
--- /dev/null
+++ b/.github/workflows/branch.yml
@@ -0,0 +1,44 @@
+name: nf-core branch protection
+# This workflow is triggered on PRs to master branch on the repository
+# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev`
+on:
+  pull_request_target:
+    branches: [master]
+
+jobs:
+  test:
+    runs-on: ubuntu-latest
+    steps:
+      # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches
+      - name: Check PRs
+        if: github.repository == 'nf-core/rnadeseq'
+        run: |
+          { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/rnadeseq ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]
+
+      # If the above check failed, post a comment on the PR explaining the failure
+      # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets
+      - name: Post PR comment
+        if: failure()
+        uses: mshick/add-pr-comment@v1
+        with:
+          message: |
+            ## This PR is against the `master` branch :x:
+
+            * Do not close this PR
+            * Click _Edit_ and change the `base` to `dev`
+            * This CI test will remain failed until you push a new commit
+
+            ---
+
+            Hi @${{ github.event.pull_request.user.login }},
+
+            It looks like this pull-request is has been made against the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `master` branch.
+            The `master` branch on nf-core repositories should always contain code from the latest release.
+            Because of this, PRs to `master` are only allowed if they come from the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `dev` branch.
+
+            You do not need to close this PR, you can change the target branch to `dev` by clicking the _"Edit"_ button at the top of this page.
+            Note that even after this, the test will continue to show as failing until you push a new commit.
+
+            Thanks again for your contribution!
+          repo-token: ${{ secrets.GITHUB_TOKEN }}
+          allow-repeats: false
diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
new file mode 100644
index 00000000..bd60b55d
--- /dev/null
+++ b/.github/workflows/ci.yml
@@ -0,0 +1,43 @@
+name: nf-core CI
+# This workflow runs the pipeline with the minimal test dataset to check that it completes without any syntax errors
+on:
+  push:
+    branches:
+      - dev
+  pull_request:
+  release:
+    types: [published]
+
+env:
+  NXF_ANSI_LOG: false
+
+concurrency:
+  group: "${{ github.workflow }}-${{ github.event.pull_request.number || github.ref }}"
+  cancel-in-progress: true
+
+jobs:
+  test:
+    name: Run pipeline with test data
+    # Only run on push if this is the nf-core dev branch (merged PRs)
+    if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/rnadeseq') }}"
+    runs-on: ubuntu-latest
+    strategy:
+      matrix:
+        NXF_VER:
+          - "22.10.1"
+          - "latest-everything"
+    steps:
+      - name: Check out pipeline code
+        uses: actions/checkout@v3
+
+      - name: Install Nextflow
+        uses: nf-core/setup-nextflow@v1
+        with:
+          version: "${{ matrix.NXF_VER }}"
+
+      - name: Run pipeline with test data
+        # TODO nf-core: You can customise CI pipeline run tests as required
+        # For example: adding multiple test runs with different parameters
+        # Remember that you can parallelise this by using strategy.matrix
+        run: |
+          nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results
diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml
new file mode 100644
index 00000000..588b4f43
--- /dev/null
+++ b/.github/workflows/fix-linting.yml
@@ -0,0 +1,55 @@
+name: Fix linting from a comment
+on:
+  issue_comment:
+    types: [created]
+
+jobs:
+  deploy:
+    # Only run if comment is on a PR with the main repo, and if it contains the magic keywords
+    if: >
+      contains(github.event.comment.html_url, '/pull/') &&
+      contains(github.event.comment.body, '@nf-core-bot fix linting') &&
+      github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      # Use the @nf-core-bot token to check out so we can push later
+      - uses: actions/checkout@v3
+        with:
+          token: ${{ secrets.nf_core_bot_auth_token }}
+
+      # Action runs on the issue comment, so we don't get the PR by default
+      # Use the gh cli to check out the PR
+      - name: Checkout Pull Request
+        run: gh pr checkout ${{ github.event.issue.number }}
+        env:
+          GITHUB_TOKEN: ${{ secrets.nf_core_bot_auth_token }}
+
+      - uses: actions/setup-node@v3
+
+      - name: Install Prettier
+        run: npm install -g prettier @prettier/plugin-php
+
+      # Check that we actually need to fix something
+      - name: Run 'prettier --check'
+        id: prettier_status
+        run: |
+          if prettier --check ${GITHUB_WORKSPACE}; then
+            echo "result=pass" >> $GITHUB_OUTPUT
+          else
+            echo "result=fail" >> $GITHUB_OUTPUT
+          fi
+
+      - name: Run 'prettier --write'
+        if: steps.prettier_status.outputs.result == 'fail'
+        run: prettier --write ${GITHUB_WORKSPACE}
+
+      - name: Commit & push changes
+        if: steps.prettier_status.outputs.result == 'fail'
+        run: |
+          git config user.email "core@nf-co.re"
+          git config user.name "nf-core-bot"
+          git config push.default upstream
+          git add .
+          git status
+          git commit -m "[automated] Fix linting with Prettier"
+          git push
diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml
new file mode 100644
index 00000000..858d622e
--- /dev/null
+++ b/.github/workflows/linting.yml
@@ -0,0 +1,108 @@
+name: nf-core linting
+# This workflow is triggered on pushes and PRs to the repository.
+# It runs the `nf-core lint` and markdown lint tests to ensure
+# that the code meets the nf-core guidelines.
+on:
+  push:
+    branches:
+      - dev
+  pull_request:
+  release:
+    types: [published]
+
+jobs:
+  EditorConfig:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - uses: actions/setup-node@v3
+
+      - name: Install editorconfig-checker
+        run: npm install -g editorconfig-checker
+
+      - name: Run ECLint check
+        run: editorconfig-checker -exclude README.md $(find .* -type f | grep -v '.git\|.py\|.md\|json\|yml\|yaml\|html\|css\|work\|.nextflow\|build\|nf_core.egg-info\|log.txt\|Makefile')
+
+  Prettier:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - uses: actions/setup-node@v3
+
+      - name: Install Prettier
+        run: npm install -g prettier
+
+      - name: Run Prettier --check
+        run: prettier --check ${GITHUB_WORKSPACE}
+
+  PythonBlack:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - name: Check code lints with Black
+        uses: psf/black@stable
+
+      # If the above check failed, post a comment on the PR explaining the failure
+      - name: Post PR comment
+        if: failure()
+        uses: mshick/add-pr-comment@v1
+        with:
+          message: |
+            ## Python linting (`black`) is failing
+
+            To keep the code consistent with lots of contributors, we run automated code consistency checks.
+            To fix this CI test, please run:
+
+            * Install [`black`](https://black.readthedocs.io/en/stable/): `pip install black`
+            * Fix formatting errors in your pipeline: `black .`
+
+            Once you push these changes the test should pass, and you can hide this comment :+1:
+
+            We highly recommend setting up Black in your code editor so that this formatting is done automatically on save. Ask about it on Slack for help!
+
+            Thanks again for your contribution!
+          repo-token: ${{ secrets.GITHUB_TOKEN }}
+          allow-repeats: false
+
+  nf-core:
+    runs-on: ubuntu-latest
+    steps:
+      - name: Check out pipeline code
+        uses: actions/checkout@v3
+
+      - name: Install Nextflow
+        uses: nf-core/setup-nextflow@v1
+
+      - uses: actions/setup-python@v4
+        with:
+          python-version: "3.7"
+          architecture: "x64"
+
+      - name: Install dependencies
+        run: |
+          python -m pip install --upgrade pip
+          pip install nf-core
+
+      - name: Run nf-core lint
+        env:
+          GITHUB_COMMENTS_URL: ${{ github.event.pull_request.comments_url }}
+          GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }}
+          GITHUB_PR_COMMIT: ${{ github.event.pull_request.head.sha }}
+        run: nf-core -l lint_log.txt lint --dir ${GITHUB_WORKSPACE} --markdown lint_results.md
+
+      - name: Save PR number
+        if: ${{ always() }}
+        run: echo ${{ github.event.pull_request.number }} > PR_number.txt
+
+      - name: Upload linting log file artifact
+        if: ${{ always() }}
+        uses: actions/upload-artifact@v3
+        with:
+          name: linting-logs
+          path: |
+            lint_log.txt
+            lint_results.md
+            PR_number.txt
diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml
new file mode 100644
index 00000000..0bbcd30f
--- /dev/null
+++ b/.github/workflows/linting_comment.yml
@@ -0,0 +1,28 @@
+name: nf-core linting comment
+# This workflow is triggered after the linting action is complete
+# It posts an automated comment to the PR, even if the PR is coming from a fork
+
+on:
+  workflow_run:
+    workflows: ["nf-core linting"]
+
+jobs:
+  test:
+    runs-on: ubuntu-latest
+    steps:
+      - name: Download lint results
+        uses: dawidd6/action-download-artifact@v2
+        with:
+          workflow: linting.yml
+          workflow_conclusion: completed
+
+      - name: Get PR number
+        id: pr_number
+        run: echo "pr_number=$(cat linting-logs/PR_number.txt)" >> $GITHUB_OUTPUT
+
+      - name: Post PR comment
+        uses: marocchino/sticky-pull-request-comment@v2
+        with:
+          GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }}
+          number: ${{ steps.pr_number.outputs.pr_number }}
+          path: linting-logs/lint_results.md
diff --git a/.gitignore b/.gitignore
new file mode 100644
index 00000000..5124c9ac
--- /dev/null
+++ b/.gitignore
@@ -0,0 +1,8 @@
+.nextflow*
+work/
+data/
+results/
+.DS_Store
+testing/
+testing*
+*.pyc
diff --git a/.gitpod.yml b/.gitpod.yml
new file mode 100644
index 00000000..85d95ecc
--- /dev/null
+++ b/.gitpod.yml
@@ -0,0 +1,14 @@
+image: nfcore/gitpod:latest
+
+vscode:
+  extensions: # based on nf-core.nf-core-extensionpack
+    - codezombiech.gitignore # Language support for .gitignore files
+    # - cssho.vscode-svgviewer                 # SVG viewer
+    - esbenp.prettier-vscode # Markdown/CommonMark linting and style checking for Visual Studio Code
+    - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed
+    - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files
+    - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar
+    - mechatroner.rainbow-csv # Highlight columns in csv files in different colors
+    # - nextflow.nextflow                      # Nextflow syntax highlighting
+    - oderwat.indent-rainbow # Highlight indentation level
+    - streetsidesoftware.code-spell-checker # Spelling checker for source code
diff --git a/.nf-core.yml b/.nf-core.yml
new file mode 100644
index 00000000..3805dc81
--- /dev/null
+++ b/.nf-core.yml
@@ -0,0 +1 @@
+repository_type: pipeline
diff --git a/.prettierignore b/.prettierignore
new file mode 100644
index 00000000..437d763d
--- /dev/null
+++ b/.prettierignore
@@ -0,0 +1,12 @@
+email_template.html
+adaptivecard.json
+slackreport.json
+.nextflow*
+work/
+data/
+results/
+.DS_Store
+testing/
+testing*
+*.pyc
+bin/
diff --git a/.prettierrc.yml b/.prettierrc.yml
new file mode 100644
index 00000000..c81f9a76
--- /dev/null
+++ b/.prettierrc.yml
@@ -0,0 +1 @@
+printWidth: 120
diff --git a/CHANGELOG.md b/CHANGELOG.md
new file mode 100644
index 00000000..9b5534b9
--- /dev/null
+++ b/CHANGELOG.md
@@ -0,0 +1,16 @@
+# nf-core/rnadeseq: Changelog
+
+The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
+and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
+
+## v2.1dev - [date]
+
+Initial release of nf-core/rnadeseq, created with the [nf-core](https://nf-co.re/) template.
+
+### `Added`
+
+### `Fixed`
+
+### `Dependencies`
+
+### `Deprecated`
diff --git a/CITATIONS.md b/CITATIONS.md
new file mode 100644
index 00000000..baaf7bea
--- /dev/null
+++ b/CITATIONS.md
@@ -0,0 +1,35 @@
+# nf-core/rnadeseq: Citations
+
+## [nf-core](https://pubmed.ncbi.nlm.nih.gov/32055031/)
+
+> Ewels PA, Peltzer A, Fillinger S, Patel H, Alneberg J, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. The nf-core framework for community-curated bioinformatics pipelines. Nat Biotechnol. 2020 Mar;38(3):276-278. doi: 10.1038/s41587-020-0439-x. PubMed PMID: 32055031.
+
+## [Nextflow](https://pubmed.ncbi.nlm.nih.gov/28398311/)
+
+> Di Tommaso P, Chatzou M, Floden EW, Barja PP, Palumbo E, Notredame C. Nextflow enables reproducible computational workflows. Nat Biotechnol. 2017 Apr 11;35(4):316-319. doi: 10.1038/nbt.3820. PubMed PMID: 28398311.
+
+## Pipeline tools
+
+- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)
+
+- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/)
+  > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924.
+
+## Software packaging/containerisation tools
+
+- [Anaconda](https://anaconda.com)
+
+  > Anaconda Software Distribution. Computer software. Vers. 2-2.4.0. Anaconda, Nov. 2016. Web.
+
+- [Bioconda](https://pubmed.ncbi.nlm.nih.gov/29967506/)
+
+  > Grüning B, Dale R, Sjödin A, Chapman BA, Rowe J, Tomkins-Tinch CH, Valieris R, Köster J; Bioconda Team. Bioconda: sustainable and comprehensive software distribution for the life sciences. Nat Methods. 2018 Jul;15(7):475-476. doi: 10.1038/s41592-018-0046-7. PubMed PMID: 29967506.
+
+- [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/)
+
+  > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671.
+
+- [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241)
+
+- [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/)
+  > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675.
diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md
new file mode 100644
index 00000000..f4fd052f
--- /dev/null
+++ b/CODE_OF_CONDUCT.md
@@ -0,0 +1,111 @@
+# Code of Conduct at nf-core (v1.0)
+
+## Our Pledge
+
+In the interest of fostering an open, collaborative, and welcoming environment, we as contributors and maintainers of nf-core, pledge to making participation in our projects and community a harassment-free experience for everyone, regardless of:
+
+- Age
+- Body size
+- Familial status
+- Gender identity and expression
+- Geographical location
+- Level of experience
+- Nationality and national origins
+- Native language
+- Physical and neurological ability
+- Race or ethnicity
+- Religion
+- Sexual identity and orientation
+- Socioeconomic status
+
+Please note that the list above is alphabetised and is therefore not ranked in any order of preference or importance.
+
+## Preamble
+
+> Note: This Code of Conduct (CoC) has been drafted by the nf-core Safety Officer and been edited after input from members of the nf-core team and others. "We", in this document, refers to the Safety Officer and members of the nf-core core team, both of whom are deemed to be members of the nf-core community and are therefore required to abide by this Code of Conduct. This document will amended periodically to keep it up-to-date, and in case of any dispute, the most current version will apply.
+
+An up-to-date list of members of the nf-core core team can be found [here](https://nf-co.re/about). Our current safety officer is Renuka Kudva.
+
+nf-core is a young and growing community that welcomes contributions from anyone with a shared vision for [Open Science Policies](https://www.fosteropenscience.eu/taxonomy/term/8). Open science policies encompass inclusive behaviours and we strive to build and maintain a safe and inclusive environment for all individuals.
+
+We have therefore adopted this code of conduct (CoC), which we require all members of our community and attendees in nf-core events to adhere to in all our workspaces at all times. Workspaces include but are not limited to Slack, meetings on Zoom, Jitsi, YouTube live etc.
+
+Our CoC will be strictly enforced and the nf-core team reserve the right to exclude participants who do not comply with our guidelines from our workspaces and future nf-core activities.
+
+We ask all members of our community to help maintain a supportive and productive workspace and to avoid behaviours that can make individuals feel unsafe or unwelcome. Please help us maintain and uphold this CoC.
+
+Questions, concerns or ideas on what we can include? Contact safety [at] nf-co [dot] re
+
+## Our Responsibilities
+
+The safety officer is responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behaviour.
+
+The safety officer in consultation with the nf-core core team have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this Code of Conduct, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful.
+
+Members of the core team or the safety officer who violate the CoC will be required to recuse themselves pending investigation. They will not have access to any reports of the violations and be subject to the same actions as others in violation of the CoC.
+
+## When are where does this Code of Conduct apply?
+
+Participation in the nf-core community is contingent on following these guidelines in all our workspaces and events. This includes but is not limited to the following listed alphabetically and therefore in no order of preference:
+
+- Communicating with an official project email address.
+- Communicating with community members within the nf-core Slack channel.
+- Participating in hackathons organised by nf-core (both online and in-person events).
+- Participating in collaborative work on GitHub, Google Suite, community calls, mentorship meetings, email correspondence.
+- Participating in workshops, training, and seminar series organised by nf-core (both online and in-person events). This applies to events hosted on web-based platforms such as Zoom, Jitsi, YouTube live etc.
+- Representing nf-core on social media. This includes both official and personal accounts.
+
+## nf-core cares 😊
+
+nf-core's CoC and expectations of respectful behaviours for all participants (including organisers and the nf-core team) include but are not limited to the following (listed in alphabetical order):
+
+- Ask for consent before sharing another community member’s personal information (including photographs) on social media.
+- Be respectful of differing viewpoints and experiences. We are all here to learn from one another and a difference in opinion can present a good learning opportunity.
+- Celebrate your accomplishments at events! (Get creative with your use of emojis 🎉 🥳 💯 🙌 !)
+- Demonstrate empathy towards other community members. (We don’t all have the same amount of time to dedicate to nf-core. If tasks are pending, don’t hesitate to gently remind members of your team. If you are leading a task, ask for help if you feel overwhelmed.)
+- Engage with and enquire after others. (This is especially important given the geographically remote nature of the nf-core community, so let’s do this the best we can)
+- Focus on what is best for the team and the community. (When in doubt, ask)
+- Graciously accept constructive criticism, yet be unafraid to question, deliberate, and learn.
+- Introduce yourself to members of the community. (We’ve all been outsiders and we know that talking to strangers can be hard for some, but remember we’re interested in getting to know you and your visions for open science!)
+- Show appreciation and **provide clear feedback**. (This is especially important because we don’t see each other in person and it can be harder to interpret subtleties. Also remember that not everyone understands a certain language to the same extent as you do, so **be clear in your communications to be kind.**)
+- Take breaks when you feel like you need them.
+- Using welcoming and inclusive language. (Participants are encouraged to display their chosen pronouns on Zoom or in communication on Slack.)
+
+## nf-core frowns on 😕
+
+The following behaviours from any participants within the nf-core community (including the organisers) will be considered unacceptable under this code of conduct. Engaging or advocating for any of the following could result in expulsion from nf-core workspaces.
+
+- Deliberate intimidation, stalking or following and sustained disruption of communication among participants of the community. This includes hijacking shared screens through actions such as using the annotate tool in conferencing software such as Zoom.
+- “Doxing” i.e. posting (or threatening to post) another person’s personal identifying information online.
+- Spamming or trolling of individuals on social media.
+- Use of sexual or discriminatory imagery, comments, or jokes and unwelcome sexual attention.
+- Verbal and text comments that reinforce social structures of domination related to gender, gender identity and expression, sexual orientation, ability, physical appearance, body size, race, age, religion or work experience.
+
+### Online Trolling
+
+The majority of nf-core interactions and events are held online. Unfortunately, holding events online comes with the added issue of online trolling. This is unacceptable, reports of such behaviour will be taken very seriously, and perpetrators will be excluded from activities immediately.
+
+All community members are required to ask members of the group they are working within for explicit consent prior to taking screenshots of individuals during video calls.
+
+## Procedures for Reporting CoC violations
+
+If someone makes you feel uncomfortable through their behaviours or actions, report it as soon as possible.
+
+You can reach out to members of the [nf-core core team](https://nf-co.re/about) and they will forward your concerns to the safety officer(s).
+
+Issues directly concerning members of the core team will be dealt with by other members of the core team and the safety manager, and possible conflicts of interest will be taken into account. nf-core is also in discussions about having an ombudsperson, and details will be shared in due course.
+
+All reports will be handled with utmost discretion and confidentially.
+
+## Attribution and Acknowledgements
+
+- The [Contributor Covenant, version 1.4](http://contributor-covenant.org/version/1/4)
+- The [OpenCon 2017 Code of Conduct](http://www.opencon2017.org/code_of_conduct) (CC BY 4.0 OpenCon organisers, SPARC and Right to Research Coalition)
+- The [eLife innovation sprint 2020 Code of Conduct](https://sprint.elifesciences.org/code-of-conduct/)
+- The [Mozilla Community Participation Guidelines v3.1](https://www.mozilla.org/en-US/about/governance/policies/participation/) (version 3.1, CC BY-SA 3.0 Mozilla)
+
+## Changelog
+
+### v1.0 - March 12th, 2021
+
+- Complete rewrite from original [Contributor Covenant](http://contributor-covenant.org/) CoC.
diff --git a/LICENSE b/LICENSE
new file mode 100644
index 00000000..b0c71392
--- /dev/null
+++ b/LICENSE
@@ -0,0 +1,21 @@
+MIT License
+
+Copyright (c) 
+
+Permission is hereby granted, free of charge, to any person obtaining a copy
+of this software and associated documentation files (the "Software"), to deal
+in the Software without restriction, including without limitation the rights
+to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
+copies of the Software, and to permit persons to whom the Software is
+furnished to do so, subject to the following conditions:
+
+The above copyright notice and this permission notice shall be included in all
+copies or substantial portions of the Software.
+
+THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
+IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
+FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE
+AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
+LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
+OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE
+SOFTWARE.
diff --git a/README.md b/README.md
new file mode 100644
index 00000000..287af5a8
--- /dev/null
+++ b/README.md
@@ -0,0 +1,92 @@
+# ![nf-core/rnadeseq](docs/images/nf-core-rnadeseq_logo_light.png#gh-light-mode-only) ![nf-core/rnadeseq](docs/images/nf-core-rnadeseq_logo_dark.png#gh-dark-mode-only)
+
+[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/rnadeseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)
+
+[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A522.10.1-23aa62.svg)](https://www.nextflow.io/)
+[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)
+[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)
+[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)
+[![Launch on Nextflow Tower](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Nextflow%20Tower-%234256e7)](https://tower.nf/launch?pipeline=https://github.com/nf-core/rnadeseq)
+
+[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23rnadeseq-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/rnadeseq)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)
+
+## Introduction
+
+<!-- TODO nf-core: Write a 1-2 sentence summary of what data the pipeline is for and what it does -->
+
+**nf-core/rnadeseq** is a bioinformatics best-practice analysis pipeline for .
+
+The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from [nf-core/modules](https://github.com/nf-core/modules) in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community!
+
+<!-- TODO nf-core: Add full-sized test dataset and amend the paragraph below if applicable -->
+
+On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources.The results obtained from the full-sized test can be viewed on the [nf-core website](https://nf-co.re/rnadeseq/results).
+
+## Pipeline summary
+
+<!-- TODO nf-core: Fill in short bullet-pointed list of the default steps in the pipeline -->
+
+1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/))
+2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))
+
+## Quick Start
+
+1. Install [`Nextflow`](https://www.nextflow.io/docs/latest/getstarted.html#installation) (`>=22.10.1`)
+
+2. Install any of [`Docker`](https://docs.docker.com/engine/installation/), [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) (you can follow [this tutorial](https://singularity-tutorial.github.io/01-installation/)), [`Podman`](https://podman.io/), [`Shifter`](https://nersc.gitlab.io/development/shifter/how-to-use/) or [`Charliecloud`](https://hpc.github.io/charliecloud/) for full pipeline reproducibility _(you can use [`Conda`](https://conda.io/miniconda.html) both to install Nextflow itself and also to manage software within pipelines. Please only use it within pipelines as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_.
+
+3. Download the pipeline and test it on a minimal dataset with a single command:
+
+   ```bash
+   nextflow run nf-core/rnadeseq -profile test,YOURPROFILE --outdir <OUTDIR>
+   ```
+
+   Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string.
+
+   > - The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`.
+   > - Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile <institute>` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment.
+   > - If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs.
+   > - If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs.
+
+4. Start running your own analysis!
+
+   <!-- TODO nf-core: Update the example "typical command" below used to run the pipeline -->
+
+   ```bash
+   nextflow run nf-core/rnadeseq --input samplesheet.csv --outdir <OUTDIR> --genome GRCh37 -profile <docker/singularity/podman/shifter/charliecloud/conda/institute>
+   ```
+
+## Documentation
+
+The nf-core/rnadeseq pipeline comes with documentation about the pipeline [usage](https://nf-co.re/rnadeseq/usage), [parameters](https://nf-co.re/rnadeseq/parameters) and [output](https://nf-co.re/rnadeseq/output).
+
+## Credits
+
+nf-core/rnadeseq was originally written by .
+
+We thank the following people for their extensive assistance in the development of this pipeline:
+
+<!-- TODO nf-core: If applicable, make list of people who have also contributed -->
+
+## Contributions and Support
+
+If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md).
+
+For further information or help, don't hesitate to get in touch on the [Slack `#rnadeseq` channel](https://nfcore.slack.com/channels/rnadeseq) (you can join with [this invite](https://nf-co.re/join/slack)).
+
+## Citations
+
+<!-- TODO nf-core: Add citation for pipeline after first release. Uncomment lines below and update Zenodo doi and badge at the top of this file. -->
+<!-- If you use  nf-core/rnadeseq for your analysis, please cite it using the following doi: [10.5281/zenodo.XXXXXX](https://doi.org/10.5281/zenodo.XXXXXX) -->
+
+<!-- TODO nf-core: Add bibliography of tools and data used in your pipeline -->
+
+An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.
+
+You can cite the `nf-core` publication as follows:
+
+> **The nf-core framework for community-curated bioinformatics pipelines.**
+>
+> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.
+>
+> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).
diff --git a/assets/adaptivecard.json b/assets/adaptivecard.json
new file mode 100644
index 00000000..99786880
--- /dev/null
+++ b/assets/adaptivecard.json
@@ -0,0 +1,67 @@
+{
+    "type": "message",
+    "attachments": [
+        {
+            "contentType": "application/vnd.microsoft.card.adaptive",
+            "contentUrl": null,
+            "content": {
+                "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json",
+                "msteams": {
+                    "width": "Full"
+                },
+                "type": "AdaptiveCard",
+                "version": "1.2",
+                "body": [
+                    {
+                        "type": "TextBlock",
+                        "size": "Large",
+                        "weight": "Bolder",
+                        "color": "<% if (success) { %>Good<% } else { %>Attention<%} %>",
+                        "text": "nf-core/rnadeseq v${version} - ${runName}",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "spacing": "None",
+                        "text": "Completed at ${dateComplete} (duration: ${duration})",
+                        "isSubtle": true,
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors. The full error message was: ${errorReport}.<% } %>",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "The command used to launch the workflow was as follows:",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "${commandLine}",
+                        "isSubtle": true,
+                        "wrap": true
+                    }
+                ],
+                "actions": [
+                    {
+                        "type": "Action.ShowCard",
+                        "title": "Pipeline Configuration",
+                        "card": {
+                            "type": "AdaptiveCard",
+                            "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json",
+                            "body": [
+                                {
+                                    "type": "FactSet",
+                                    "facts": [<% out << summary.collect{ k,v -> "{\"title\": \"$k\", \"value\" : \"$v\"}"}.join(",\n") %>
+                                    ]
+                                }
+                            ]
+                        }
+                    }
+                ]
+            }
+        }
+    ]
+}
diff --git a/assets/email_template.html b/assets/email_template.html
new file mode 100644
index 00000000..cffc0892
--- /dev/null
+++ b/assets/email_template.html
@@ -0,0 +1,53 @@
+<html>
+<head>
+  <meta charset="utf-8">
+  <meta http-equiv="X-UA-Compatible" content="IE=edge">
+  <meta name="viewport" content="width=device-width, initial-scale=1">
+
+  <meta name="description" content="nf-core/rnadeseq: ">
+  <title>nf-core/rnadeseq Pipeline Report</title>
+</head>
+<body>
+<div style="font-family: Helvetica, Arial, sans-serif; padding: 30px; max-width: 800px; margin: 0 auto;">
+
+<img src="cid:nfcorepipelinelogo">
+
+<h1>nf-core/rnadeseq v${version}</h1>
+<h2>Run Name: $runName</h2>
+
+<% if (!success){
+    out << """
+    <div style="color: #a94442; background-color: #f2dede; border-color: #ebccd1; padding: 15px; margin-bottom: 20px; border: 1px solid transparent; border-radius: 4px;">
+        <h4 style="margin-top:0; color: inherit;">nf-core/rnadeseq execution completed unsuccessfully!</h4>
+        <p>The exit status of the task that caused the workflow execution to fail was: <code>$exitStatus</code>.</p>
+        <p>The full error message was:</p>
+        <pre style="white-space: pre-wrap; overflow: visible; margin-bottom: 0;">${errorReport}</pre>
+    </div>
+    """
+} else {
+    out << """
+    <div style="color: #3c763d; background-color: #dff0d8; border-color: #d6e9c6; padding: 15px; margin-bottom: 20px; border: 1px solid transparent; border-radius: 4px;">
+        nf-core/rnadeseq execution completed successfully!
+    </div>
+    """
+}
+%>
+
+<p>The workflow was completed at <strong>$dateComplete</strong> (duration: <strong>$duration</strong>)</p>
+<p>The command used to launch the workflow was as follows:</p>
+<pre style="white-space: pre-wrap; overflow: visible; background-color: #ededed; padding: 15px; border-radius: 4px; margin-bottom:30px;">$commandLine</pre>
+
+<h3>Pipeline Configuration:</h3>
+<table style="width:100%; max-width:100%; border-spacing: 0; border-collapse: collapse; border:0; margin-bottom: 30px;">
+    <tbody style="border-bottom: 1px solid #ddd;">
+        <% out << summary.collect{ k,v -> "<tr><th style='text-align:left; padding: 8px 0; line-height: 1.42857143; vertical-align: top; border-top: 1px solid #ddd;'>$k</th><td style='text-align:left; padding: 8px; line-height: 1.42857143; vertical-align: top; border-top: 1px solid #ddd;'><pre style='white-space: pre-wrap; overflow: visible;'>$v</pre></td></tr>" }.join("\n") %>
+    </tbody>
+</table>
+
+<p>nf-core/rnadeseq</p>
+<p><a href="https://github.com/nf-core/rnadeseq">https://github.com/nf-core/rnadeseq</a></p>
+
+</div>
+
+</body>
+</html>
diff --git a/assets/email_template.txt b/assets/email_template.txt
new file mode 100644
index 00000000..1a48efa9
--- /dev/null
+++ b/assets/email_template.txt
@@ -0,0 +1,39 @@
+----------------------------------------------------
+                                        ,--./,-.
+        ___     __   __   __   ___     /,-._.--~\\
+  |\\ | |__  __ /  ` /  \\ |__) |__         }  {
+  | \\| |       \\__, \\__/ |  \\ |___     \\`-._,-`-,
+                                        `._,._,'
+  nf-core/rnadeseq v${version}
+----------------------------------------------------
+Run Name: $runName
+
+<% if (success){
+    out << "## nf-core/rnadeseq execution completed successfully! ##"
+} else {
+    out << """####################################################
+## nf-core/rnadeseq execution completed unsuccessfully! ##
+####################################################
+The exit status of the task that caused the workflow execution to fail was: $exitStatus.
+The full error message was:
+
+${errorReport}
+"""
+} %>
+
+
+The workflow was completed at $dateComplete (duration: $duration)
+
+The command used to launch the workflow was as follows:
+
+  $commandLine
+
+
+
+Pipeline Configuration:
+-----------------------
+<% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %>
+
+--
+nf-core/rnadeseq
+https://github.com/nf-core/rnadeseq
diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml
new file mode 100644
index 00000000..29a63bbb
--- /dev/null
+++ b/assets/methods_description_template.yml
@@ -0,0 +1,25 @@
+id: "nf-core-rnadeseq-methods-description"
+description: "Suggested text and references to use when describing pipeline usage within the methods section of a publication."
+section_name: "nf-core/rnadeseq Methods Description"
+section_href: "https://github.com/nf-core/rnadeseq"
+plot_type: "html"
+## TODO nf-core: Update the HTML below to your prefered methods description, e.g. add publication citation for this pipeline
+## You inject any metadata in the Nextflow '${workflow}' object
+data: |
+  <h4>Methods</h4>
+  <p>Data was processed using nf-core/rnadeseq v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (<a href="https://doi.org/10.1038/s41587-020-0439-x">Ewels <em>et al.</em>, 2020</a>).</p>
+  <p>The pipeline was executed with Nextflow v${workflow.nextflow.version} (<a href="https://doi.org/10.1038/nbt.3820">Di Tommaso <em>et al.</em>, 2017</a>) with the following command:</p>
+  <pre><code>${workflow.commandLine}</code></pre>
+  <h4>References</h4>
+  <ul>
+    <li>Di Tommaso, P., Chatzou, M., Floden, E. W., Barja, P. P., Palumbo, E., & Notredame, C. (2017). Nextflow enables reproducible computational workflows. Nature Biotechnology, 35(4), 316-319. <a href="https://doi.org/10.1038/nbt.3820">https://doi.org/10.1038/nbt.3820</a></li>
+    <li>Ewels, P. A., Peltzer, A., Fillinger, S., Patel, H., Alneberg, J., Wilm, A., Garcia, M. U., Di Tommaso, P., & Nahnsen, S. (2020). The nf-core framework for community-curated bioinformatics pipelines. Nature Biotechnology, 38(3), 276-278. <a href="https://doi.org/10.1038/s41587-020-0439-x">https://doi.org/10.1038/s41587-020-0439-x</a></li>
+  </ul>
+  <div class="alert alert-info">
+    <h5>Notes:</h5>
+    <ul>
+      ${nodoi_text}
+      <li>The command above does not include parameters contained in any configs or profiles that may have been used. Ensure the config file is also uploaded with your publication!</li>
+      <li>You should also cite all software used within this run. Check the "Software Versions" of this report to get version information.</li>
+    </ul>
+  </div>
diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
new file mode 100644
index 00000000..8d198f08
--- /dev/null
+++ b/assets/multiqc_config.yml
@@ -0,0 +1,13 @@
+report_comment: >
+  This report has been generated by the <a href="https://github.com/nf-core/rnadeseq" target="_blank">nf-core/rnadeseq</a>
+  analysis pipeline. For information about how to interpret these results, please see the
+  <a href="https://nf-co.re/rnadeseq" target="_blank">documentation</a>.
+report_section_order:
+  "nf-core-rnadeseq-methods-description":
+    order: -1000
+  software_versions:
+    order: -1001
+  "nf-core-rnadeseq-summary":
+    order: -1002
+
+export_plots: true
diff --git a/assets/nf-core-rnadeseq_logo_light.png b/assets/nf-core-rnadeseq_logo_light.png
new file mode 100644
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diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv
new file mode 100644
index 00000000..5f653ab7
--- /dev/null
+++ b/assets/samplesheet.csv
@@ -0,0 +1,3 @@
+sample,fastq_1,fastq_2
+SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz
+SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz,
diff --git a/assets/schema_input.json b/assets/schema_input.json
new file mode 100644
index 00000000..bd54c145
--- /dev/null
+++ b/assets/schema_input.json
@@ -0,0 +1,36 @@
+{
+    "$schema": "http://json-schema.org/draft-07/schema",
+    "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/assets/schema_input.json",
+    "title": "nf-core/rnadeseq pipeline - params.input schema",
+    "description": "Schema for the file provided with params.input",
+    "type": "array",
+    "items": {
+        "type": "object",
+        "properties": {
+            "sample": {
+                "type": "string",
+                "pattern": "^\\S+$",
+                "errorMessage": "Sample name must be provided and cannot contain spaces"
+            },
+            "fastq_1": {
+                "type": "string",
+                "pattern": "^\\S+\\.f(ast)?q\\.gz$",
+                "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'"
+            },
+            "fastq_2": {
+                "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'",
+                "anyOf": [
+                    {
+                        "type": "string",
+                        "pattern": "^\\S+\\.f(ast)?q\\.gz$"
+                    },
+                    {
+                        "type": "string",
+                        "maxLength": 0
+                    }
+                ]
+            }
+        },
+        "required": ["sample", "fastq_1"]
+    }
+}
diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt
new file mode 100644
index 00000000..953c8470
--- /dev/null
+++ b/assets/sendmail_template.txt
@@ -0,0 +1,53 @@
+To: $email
+Subject: $subject
+Mime-Version: 1.0
+Content-Type: multipart/related;boundary="nfcoremimeboundary"
+
+--nfcoremimeboundary
+Content-Type: text/html; charset=utf-8
+
+$email_html
+
+--nfcoremimeboundary
+Content-Type: image/png;name="nf-core-rnadeseq_logo.png"
+Content-Transfer-Encoding: base64
+Content-ID: <nfcorepipelinelogo>
+Content-Disposition: inline; filename="nf-core-rnadeseq_logo_light.png"
+
+<% out << new File("$projectDir/assets/nf-core-rnadeseq_logo_light.png").
+    bytes.
+    encodeBase64().
+    toString().
+    tokenize( '\n' )*.
+    toList()*.
+    collate( 76 )*.
+    collect { it.join() }.
+    flatten().
+    join( '\n' ) %>
+
+<%
+if (mqcFile){
+def mqcFileObj = new File("$mqcFile")
+if (mqcFileObj.length() < mqcMaxSize){
+out << """
+--nfcoremimeboundary
+Content-Type: text/html; name=\"multiqc_report\"
+Content-Transfer-Encoding: base64
+Content-ID: <mqcreport>
+Content-Disposition: attachment; filename=\"${mqcFileObj.getName()}\"
+
+${mqcFileObj.
+    bytes.
+    encodeBase64().
+    toString().
+    tokenize( '\n' )*.
+    toList()*.
+    collate( 76 )*.
+    collect { it.join() }.
+    flatten().
+    join( '\n' )}
+"""
+}}
+%>
+
+--nfcoremimeboundary--
diff --git a/assets/slackreport.json b/assets/slackreport.json
new file mode 100644
index 00000000..043d02f2
--- /dev/null
+++ b/assets/slackreport.json
@@ -0,0 +1,34 @@
+{
+    "attachments": [
+        {
+            "fallback": "Plain-text summary of the attachment.",
+            "color": "<% if (success) { %>good<% } else { %>danger<%} %>",
+            "author_name": "sanger-tol/readmapping v${version} - ${runName}",
+            "author_icon": "https://www.nextflow.io/docs/latest/_static/favicon.ico",
+            "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors<% } %>",
+            "fields": [
+                {
+                    "title": "Command used to launch the workflow",
+                    "value": "```${commandLine}```",
+                    "short": false
+                }
+                <%
+                    if (!success) { %>
+                    ,
+                    {
+                        "title": "Full error message",
+                        "value": "```${errorReport}```",
+                        "short": false
+                    },
+                    {
+                        "title": "Pipeline configuration",
+                        "value": "<% out << summary.collect{ k,v -> k == "hook_url" ? "_${k}_: (_hidden_)" : ( ( v.class.toString().contains('Path') || ( v.class.toString().contains('String') && v.contains('/') ) ) ? "_${k}_: `${v}`" : (v.class.toString().contains('DateTime') ? ("_${k}_: " + v.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM))) : "_${k}_: ${v}") ) }.join(",\n") %>",
+                        "short": false
+                    }
+                    <% }
+                %>
+            ],
+            "footer": "Completed at <% out << dateComplete.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM)) %> (duration: ${duration})"
+        }
+    ]
+}
diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py
new file mode 100755
index 00000000..11b15572
--- /dev/null
+++ b/bin/check_samplesheet.py
@@ -0,0 +1,262 @@
+#!/usr/bin/env python
+
+
+"""Provide a command line tool to validate and transform tabular samplesheets."""
+
+
+import argparse
+import csv
+import logging
+import sys
+from collections import Counter
+from pathlib import Path
+
+logger = logging.getLogger()
+
+
+class RowChecker:
+    """
+    Define a service that can validate and transform each given row.
+
+    Attributes:
+        modified (list): A list of dicts, where each dict corresponds to a previously
+            validated and transformed row. The order of rows is maintained.
+
+    """
+
+    VALID_FORMATS = (
+        ".fq.gz",
+        ".fastq.gz",
+    )
+
+    def __init__(
+        self,
+        sample_col="sample",
+        first_col="fastq_1",
+        second_col="fastq_2",
+        single_col="single_end",
+        **kwargs,
+    ):
+        """
+        Initialize the row checker with the expected column names.
+
+        Args:
+            sample_col (str): The name of the column that contains the sample name
+                (default "sample").
+            first_col (str): The name of the column that contains the first (or only)
+                FASTQ file path (default "fastq_1").
+            second_col (str): The name of the column that contains the second (if any)
+                FASTQ file path (default "fastq_2").
+            single_col (str): The name of the new column that will be inserted and
+                records whether the sample contains single- or paired-end sequencing
+                reads (default "single_end").
+
+        """
+        super().__init__(**kwargs)
+        self._sample_col = sample_col
+        self._first_col = first_col
+        self._second_col = second_col
+        self._single_col = single_col
+        self._seen = set()
+        self.modified = []
+
+    def validate_and_transform(self, row):
+        """
+        Perform all validations on the given row and insert the read pairing status.
+
+        Args:
+            row (dict): A mapping from column headers (keys) to elements of that row
+                (values).
+
+        """
+        self._validate_sample(row)
+        self._validate_first(row)
+        self._validate_second(row)
+        self._validate_pair(row)
+        self._seen.add((row[self._sample_col], row[self._first_col]))
+        self.modified.append(row)
+
+    def _validate_sample(self, row):
+        """Assert that the sample name exists and convert spaces to underscores."""
+        if len(row[self._sample_col]) <= 0:
+            raise AssertionError("Sample input is required.")
+        # Sanitize samples slightly.
+        row[self._sample_col] = row[self._sample_col].replace(" ", "_")
+
+    def _validate_first(self, row):
+        """Assert that the first FASTQ entry is non-empty and has the right format."""
+        if len(row[self._first_col]) <= 0:
+            raise AssertionError("At least the first FASTQ file is required.")
+        self._validate_fastq_format(row[self._first_col])
+
+    def _validate_second(self, row):
+        """Assert that the second FASTQ entry has the right format if it exists."""
+        if len(row[self._second_col]) > 0:
+            self._validate_fastq_format(row[self._second_col])
+
+    def _validate_pair(self, row):
+        """Assert that read pairs have the same file extension. Report pair status."""
+        if row[self._first_col] and row[self._second_col]:
+            row[self._single_col] = False
+            first_col_suffix = Path(row[self._first_col]).suffixes[-2:]
+            second_col_suffix = Path(row[self._second_col]).suffixes[-2:]
+            if first_col_suffix != second_col_suffix:
+                raise AssertionError("FASTQ pairs must have the same file extensions.")
+        else:
+            row[self._single_col] = True
+
+    def _validate_fastq_format(self, filename):
+        """Assert that a given filename has one of the expected FASTQ extensions."""
+        if not any(filename.endswith(extension) for extension in self.VALID_FORMATS):
+            raise AssertionError(
+                f"The FASTQ file has an unrecognized extension: {filename}\n"
+                f"It should be one of: {', '.join(self.VALID_FORMATS)}"
+            )
+
+    def validate_unique_samples(self):
+        """
+        Assert that the combination of sample name and FASTQ filename is unique.
+
+        In addition to the validation, also rename all samples to have a suffix of _T{n}, where n is the
+        number of times the same sample exist, but with different FASTQ files, e.g., multiple runs per experiment.
+
+        """
+        if len(self._seen) != len(self.modified):
+            raise AssertionError("The pair of sample name and FASTQ must be unique.")
+        seen = Counter()
+        for row in self.modified:
+            sample = row[self._sample_col]
+            seen[sample] += 1
+            row[self._sample_col] = f"{sample}_T{seen[sample]}"
+
+
+def read_head(handle, num_lines=10):
+    """Read the specified number of lines from the current position in the file."""
+    lines = []
+    for idx, line in enumerate(handle):
+        if idx == num_lines:
+            break
+        lines.append(line)
+    return "".join(lines)
+
+
+def sniff_format(handle):
+    """
+    Detect the tabular format.
+
+    Args:
+        handle (text file): A handle to a `text file`_ object. The read position is
+        expected to be at the beginning (index 0).
+
+    Returns:
+        csv.Dialect: The detected tabular format.
+
+    .. _text file:
+        https://docs.python.org/3/glossary.html#term-text-file
+
+    """
+    peek = read_head(handle)
+    handle.seek(0)
+    sniffer = csv.Sniffer()
+    if not sniffer.has_header(peek):
+        logger.critical("The given sample sheet does not appear to contain a header.")
+        sys.exit(1)
+    dialect = sniffer.sniff(peek)
+    return dialect
+
+
+def check_samplesheet(file_in, file_out):
+    """
+    Check that the tabular samplesheet has the structure expected by nf-core pipelines.
+
+    Validate the general shape of the table, expected columns, and each row. Also add
+    an additional column which records whether one or two FASTQ reads were found.
+
+    Args:
+        file_in (pathlib.Path): The given tabular samplesheet. The format can be either
+            CSV, TSV, or any other format automatically recognized by ``csv.Sniffer``.
+        file_out (pathlib.Path): Where the validated and transformed samplesheet should
+            be created; always in CSV format.
+
+    Example:
+        This function checks that the samplesheet follows the following structure,
+        see also the `viral recon samplesheet`_::
+
+            sample,fastq_1,fastq_2
+            SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz
+            SAMPLE_PE,SAMPLE_PE_RUN2_1.fastq.gz,SAMPLE_PE_RUN2_2.fastq.gz
+            SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz,
+
+    .. _viral recon samplesheet:
+        https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv
+
+    """
+    required_columns = {"sample", "fastq_1", "fastq_2"}
+    # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`.
+    with file_in.open(newline="") as in_handle:
+        reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle))
+        # Validate the existence of the expected header columns.
+        if not required_columns.issubset(reader.fieldnames):
+            req_cols = ", ".join(required_columns)
+            logger.critical(f"The sample sheet **must** contain these column headers: {req_cols}.")
+            sys.exit(1)
+        # Validate each row.
+        checker = RowChecker()
+        for i, row in enumerate(reader):
+            try:
+                checker.validate_and_transform(row)
+            except AssertionError as error:
+                logger.critical(f"{str(error)} On line {i + 2}.")
+                sys.exit(1)
+        checker.validate_unique_samples()
+    header = list(reader.fieldnames)
+    header.insert(1, "single_end")
+    # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`.
+    with file_out.open(mode="w", newline="") as out_handle:
+        writer = csv.DictWriter(out_handle, header, delimiter=",")
+        writer.writeheader()
+        for row in checker.modified:
+            writer.writerow(row)
+
+
+def parse_args(argv=None):
+    """Define and immediately parse command line arguments."""
+    parser = argparse.ArgumentParser(
+        description="Validate and transform a tabular samplesheet.",
+        epilog="Example: python check_samplesheet.py samplesheet.csv samplesheet.valid.csv",
+    )
+    parser.add_argument(
+        "file_in",
+        metavar="FILE_IN",
+        type=Path,
+        help="Tabular input samplesheet in CSV or TSV format.",
+    )
+    parser.add_argument(
+        "file_out",
+        metavar="FILE_OUT",
+        type=Path,
+        help="Transformed output samplesheet in CSV format.",
+    )
+    parser.add_argument(
+        "-l",
+        "--log-level",
+        help="The desired log level (default WARNING).",
+        choices=("CRITICAL", "ERROR", "WARNING", "INFO", "DEBUG"),
+        default="WARNING",
+    )
+    return parser.parse_args(argv)
+
+
+def main(argv=None):
+    """Coordinate argument parsing and program execution."""
+    args = parse_args(argv)
+    logging.basicConfig(level=args.log_level, format="[%(levelname)s] %(message)s")
+    if not args.file_in.is_file():
+        logger.error(f"The given input file {args.file_in} was not found!")
+        sys.exit(2)
+    args.file_out.parent.mkdir(parents=True, exist_ok=True)
+    check_samplesheet(args.file_in, args.file_out)
+
+
+if __name__ == "__main__":
+    sys.exit(main())
diff --git a/conf/base.config b/conf/base.config
new file mode 100644
index 00000000..ff68a002
--- /dev/null
+++ b/conf/base.config
@@ -0,0 +1,65 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq Nextflow base config file
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    A 'blank slate' config file, appropriate for general use on most high performance
+    compute environments. Assumes that all software is installed and available on
+    the PATH. Runs in `local` mode - all jobs will be run on the logged in environment.
+----------------------------------------------------------------------------------------
+*/
+
+process {
+
+    // TODO nf-core: Check the defaults for all processes
+    cpus   = { check_max( 1    * task.attempt, 'cpus'   ) }
+    memory = { check_max( 6.GB * task.attempt, 'memory' ) }
+    time   = { check_max( 4.h  * task.attempt, 'time'   ) }
+
+    errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' }
+    maxRetries    = 1
+    maxErrors     = '-1'
+
+    // Process-specific resource requirements
+    // NOTE - Please try and re-use the labels below as much as possible.
+    //        These labels are used and recognised by default in DSL2 files hosted on nf-core/modules.
+    //        If possible, it would be nice to keep the same label naming convention when
+    //        adding in your local modules too.
+    // TODO nf-core: Customise requirements for specific processes.
+    // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors
+    withLabel:process_single {
+        cpus   = { check_max( 1                  , 'cpus'    ) }
+        memory = { check_max( 6.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 4.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_low {
+        cpus   = { check_max( 2     * task.attempt, 'cpus'    ) }
+        memory = { check_max( 12.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 4.h   * task.attempt, 'time'    ) }
+    }
+    withLabel:process_medium {
+        cpus   = { check_max( 6     * task.attempt, 'cpus'    ) }
+        memory = { check_max( 36.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 8.h   * task.attempt, 'time'    ) }
+    }
+    withLabel:process_high {
+        cpus   = { check_max( 12    * task.attempt, 'cpus'    ) }
+        memory = { check_max( 72.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 16.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_long {
+        time   = { check_max( 20.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_high_memory {
+        memory = { check_max( 200.GB * task.attempt, 'memory' ) }
+    }
+    withLabel:error_ignore {
+        errorStrategy = 'ignore'
+    }
+    withLabel:error_retry {
+        errorStrategy = 'retry'
+        maxRetries    = 2
+    }
+    withName:CUSTOM_DUMPSOFTWAREVERSIONS {
+        cache = false
+    }
+}
diff --git a/conf/igenomes.config b/conf/igenomes.config
new file mode 100644
index 00000000..7a1b3ac6
--- /dev/null
+++ b/conf/igenomes.config
@@ -0,0 +1,432 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for iGenomes paths
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines reference genomes using iGenome paths.
+    Can be used by any config that customises the base path using:
+        $params.igenomes_base / --igenomes_base
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    // illumina iGenomes reference file paths
+    genomes {
+        'GRCh37' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/GRCh37-blacklist.bed"
+        }
+        'GRCh38' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg38-blacklist.bed"
+        }
+        'GRCm38' {
+            fasta       = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "1.87e9"
+            blacklist   = "${projectDir}/assets/blacklists/GRCm38-blacklist.bed"
+        }
+        'TAIR10' {
+            fasta       = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/README.txt"
+            mito_name   = "Mt"
+        }
+        'EB2' {
+            fasta       = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/README.txt"
+        }
+        'UMD3.1' {
+            fasta       = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'WBcel235' {
+            fasta       = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed"
+            mito_name   = "MtDNA"
+            macs_gsize  = "9e7"
+        }
+        'CanFam3.1' {
+            fasta       = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'GRCz10' {
+            fasta       = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'BDGP6' {
+            fasta       = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed"
+            mito_name   = "M"
+            macs_gsize  = "1.2e8"
+        }
+        'EquCab2' {
+            fasta       = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'EB1' {
+            fasta       = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/README.txt"
+        }
+        'Galgal4' {
+            fasta       = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'Gm01' {
+            fasta       = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/README.txt"
+        }
+        'Mmul_1' {
+            fasta       = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'IRGSP-1.0' {
+            fasta       = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed"
+            mito_name   = "Mt"
+        }
+        'CHIMP2.1.4' {
+            fasta       = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'Rnor_5.0' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'Rnor_6.0' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'R64-1-1' {
+            fasta       = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+            macs_gsize  = "1.2e7"
+        }
+        'EF2' {
+            fasta       = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "1.21e7"
+        }
+        'Sbi1' {
+            fasta       = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/README.txt"
+        }
+        'Sscrofa10.2' {
+            fasta       = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'AGPv3' {
+            fasta       = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed"
+            mito_name   = "Mt"
+        }
+        'hg38' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg38-blacklist.bed"
+        }
+        'hg19' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg19-blacklist.bed"
+        }
+        'mm10' {
+            fasta       = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "1.87e9"
+            blacklist   = "${projectDir}/assets/blacklists/mm10-blacklist.bed"
+        }
+        'bosTau8' {
+            fasta       = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+        }
+        'ce10' {
+            fasta       = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "9e7"
+        }
+        'canFam3' {
+            fasta       = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'danRer10' {
+            fasta       = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "1.37e9"
+        }
+        'dm6' {
+            fasta       = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "1.2e8"
+        }
+        'equCab2' {
+            fasta       = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'galGal4' {
+            fasta       = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'panTro4' {
+            fasta       = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'rn6' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+        }
+        'sacCer3' {
+            fasta       = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/"
+            readme      = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "1.2e7"
+        }
+        'susScr3' {
+            fasta       = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+    }
+}
diff --git a/conf/modules.config b/conf/modules.config
new file mode 100644
index 00000000..da58a5d8
--- /dev/null
+++ b/conf/modules.config
@@ -0,0 +1,41 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Config file for defining DSL2 per module options and publishing paths
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Available keys to override module options:
+        ext.args   = Additional arguments appended to command in module.
+        ext.args2  = Second set of arguments appended to command in module (multi-tool modules).
+        ext.args3  = Third set of arguments appended to command in module (multi-tool modules).
+        ext.prefix = File name prefix for output files.
+----------------------------------------------------------------------------------------
+*/
+
+process {
+
+    publishDir = [
+        path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" },
+        mode: params.publish_dir_mode,
+        saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+    ]
+
+    withName: SAMPLESHEET_CHECK {
+        publishDir = [
+            path: { "${params.outdir}/pipeline_info" },
+            mode: params.publish_dir_mode,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: FASTQC {
+        ext.args = '--quiet'
+    }
+
+    withName: CUSTOM_DUMPSOFTWAREVERSIONS {
+        publishDir = [
+            path: { "${params.outdir}/pipeline_info" },
+            mode: params.publish_dir_mode,
+            pattern: '*_versions.yml'
+        ]
+    }
+
+}
diff --git a/conf/test.config b/conf/test.config
new file mode 100644
index 00000000..918ce810
--- /dev/null
+++ b/conf/test.config
@@ -0,0 +1,29 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for running minimal tests
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines input files and everything required to run a fast and simple pipeline test.
+
+    Use as follows:
+        nextflow run nf-core/rnadeseq -profile test,<docker/singularity> --outdir <OUTDIR>
+
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    config_profile_name        = 'Test profile'
+    config_profile_description = 'Minimal test dataset to check pipeline function'
+
+    // Limit resources so that this can run on GitHub Actions
+    max_cpus   = 2
+    max_memory = '6.GB'
+    max_time   = '6.h'
+
+    // Input data
+    // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets
+    // TODO nf-core: Give any required params for the test so that command line flags are not needed
+    input  = 'https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv'
+
+    // Genome references
+    genome = 'R64-1-1'
+}
diff --git a/conf/test_full.config b/conf/test_full.config
new file mode 100644
index 00000000..126f6d5a
--- /dev/null
+++ b/conf/test_full.config
@@ -0,0 +1,24 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for running full-size tests
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines input files and everything required to run a full size pipeline test.
+
+    Use as follows:
+        nextflow run nf-core/rnadeseq -profile test_full,<docker/singularity> --outdir <OUTDIR>
+
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    config_profile_name        = 'Full test profile'
+    config_profile_description = 'Full test dataset to check pipeline function'
+
+    // Input data for full size test
+    // TODO nf-core: Specify the paths to your full test data ( on nf-core/test-datasets or directly in repositories, e.g. SRA)
+    // TODO nf-core: Give any required params for the test so that command line flags are not needed
+    input = 'https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_full_illumina_amplicon.csv'
+
+    // Genome references
+    genome = 'R64-1-1'
+}
diff --git a/docs/README.md b/docs/README.md
new file mode 100644
index 00000000..dc052cc1
--- /dev/null
+++ b/docs/README.md
@@ -0,0 +1,10 @@
+# nf-core/rnadeseq: Documentation
+
+The nf-core/rnadeseq documentation is split into the following pages:
+
+- [Usage](usage.md)
+  - An overview of how the pipeline works, how to run it and a description of all of the different command-line flags.
+- [Output](output.md)
+  - An overview of the different results produced by the pipeline and how to interpret them.
+
+You can find a lot more documentation about installing, configuring and running nf-core pipelines on the website: [https://nf-co.re](https://nf-co.re)
diff --git a/docs/images/mqc_fastqc_adapter.png b/docs/images/mqc_fastqc_adapter.png
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diff --git a/docs/output.md b/docs/output.md
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index 00000000..34c44ec1
--- /dev/null
+++ b/docs/output.md
@@ -0,0 +1,68 @@
+# nf-core/rnadeseq: Output
+
+## Introduction
+
+This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline.
+
+The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory.
+
+<!-- TODO nf-core: Write this documentation describing your workflow's output -->
+
+## Pipeline overview
+
+The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps:
+
+- [FastQC](#fastqc) - Raw read QC
+- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline
+- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution
+
+### FastQC
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `fastqc/`
+  - `*_fastqc.html`: FastQC report containing quality metrics.
+  - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images.
+
+</details>
+
+[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your sequenced reads. It provides information about the quality score distribution across your reads, per base sequence content (%A/T/G/C), adapter contamination and overrepresented sequences. For further reading and documentation see the [FastQC help pages](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/).
+
+![MultiQC - FastQC sequence counts plot](images/mqc_fastqc_counts.png)
+
+![MultiQC - FastQC mean quality scores plot](images/mqc_fastqc_quality.png)
+
+![MultiQC - FastQC adapter content plot](images/mqc_fastqc_adapter.png)
+
+> **NB:** The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality.
+
+### MultiQC
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `multiqc/`
+  - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
+  - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
+  - `multiqc_plots/`: directory containing static images from the report in various formats.
+
+</details>
+
+[MultiQC](http://multiqc.info) is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory.
+
+Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see <http://multiqc.info>.
+
+### Pipeline information
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `pipeline_info/`
+  - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`.
+  - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline.
+  - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`.
+
+</details>
+
+[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to troubleshoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage.
diff --git a/docs/usage.md b/docs/usage.md
new file mode 100644
index 00000000..4a6d51e8
--- /dev/null
+++ b/docs/usage.md
@@ -0,0 +1,270 @@
+# nf-core/rnadeseq: Usage
+
+## :warning: Please read this documentation on the nf-core website: [https://nf-co.re/rnadeseq/usage](https://nf-co.re/rnadeseq/usage)
+
+> _Documentation of pipeline parameters is generated automatically from the pipeline schema and can no longer be found in markdown files._
+
+## Introduction
+
+<!-- TODO nf-core: Add documentation about anything specific to running your pipeline. For general topics, please point to (and add to) the main nf-core website. -->
+
+## Samplesheet input
+
+You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below.
+
+```bash
+--input '[path to samplesheet file]'
+```
+
+### Multiple runs of the same sample
+
+The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. The pipeline will concatenate the raw reads before performing any downstream analysis. Below is an example for the same sample sequenced across 3 lanes:
+
+```console
+sample,fastq_1,fastq_2
+CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
+CONTROL_REP1,AEG588A1_S1_L003_R1_001.fastq.gz,AEG588A1_S1_L003_R2_001.fastq.gz
+CONTROL_REP1,AEG588A1_S1_L004_R1_001.fastq.gz,AEG588A1_S1_L004_R2_001.fastq.gz
+```
+
+### Full samplesheet
+
+The pipeline will auto-detect whether a sample is single- or paired-end using the information provided in the samplesheet. The samplesheet can have as many columns as you desire, however, there is a strict requirement for the first 3 columns to match those defined in the table below.
+
+A final samplesheet file consisting of both single- and paired-end data may look something like the one below. This is for 6 samples, where `TREATMENT_REP3` has been sequenced twice.
+
+```console
+sample,fastq_1,fastq_2
+CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
+CONTROL_REP2,AEG588A2_S2_L002_R1_001.fastq.gz,AEG588A2_S2_L002_R2_001.fastq.gz
+CONTROL_REP3,AEG588A3_S3_L002_R1_001.fastq.gz,AEG588A3_S3_L002_R2_001.fastq.gz
+TREATMENT_REP1,AEG588A4_S4_L003_R1_001.fastq.gz,
+TREATMENT_REP2,AEG588A5_S5_L003_R1_001.fastq.gz,
+TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz,
+TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz,
+```
+
+| Column    | Description                                                                                                                                                                            |
+| --------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
+| `sample`  | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). |
+| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz".                                                             |
+| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz".                                                             |
+
+An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline.
+
+## Running the pipeline
+
+The typical command for running the pipeline is as follows:
+
+```bash
+nextflow run nf-core/rnadeseq --input samplesheet.csv --outdir <OUTDIR> --genome GRCh37 -profile docker
+```
+
+This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles.
+
+Note that the pipeline will create the following files in your working directory:
+
+```bash
+work                # Directory containing the nextflow working files
+<OUTDIR>            # Finished results in specified location (defined with --outdir)
+.nextflow_log       # Log file from Nextflow
+# Other nextflow hidden files, eg. history of pipeline runs and old logs.
+```
+
+### Updating the pipeline
+
+When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline:
+
+```bash
+nextflow pull nf-core/rnadeseq
+```
+
+### Reproducibility
+
+It is a good idea to specify a pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since.
+
+First, go to the [nf-core/rnadeseq releases page](https://github.com/nf-core/rnadeseq/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag.
+
+This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. For example, at the bottom of the MultiQC reports.
+
+## Core Nextflow arguments
+
+> **NB:** These options are part of Nextflow and use a _single_ hyphen (pipeline parameters use a double-hyphen).
+
+### `-profile`
+
+Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments.
+
+Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Podman, Shifter, Charliecloud, Conda) - see below.
+
+> We highly recommend the use of Docker or Singularity containers for full pipeline reproducibility, however when this is not possible, Conda is also supported.
+
+The pipeline also dynamically loads configurations from [https://github.com/nf-core/configs](https://github.com/nf-core/configs) when it runs, making multiple config profiles for various institutional clusters available at run time. For more information and to see if your system is available in these configs please see the [nf-core/configs documentation](https://github.com/nf-core/configs#documentation).
+
+Note that multiple profiles can be loaded, for example: `-profile test,docker` - the order of arguments is important!
+They are loaded in sequence, so later profiles can overwrite earlier profiles.
+
+If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended, since it can lead to different results on different machines dependent on the computer enviroment.
+
+- `test`
+  - A profile with a complete configuration for automated testing
+  - Includes links to test data so needs no other parameters
+- `docker`
+  - A generic configuration profile to be used with [Docker](https://docker.com/)
+- `singularity`
+  - A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/)
+- `podman`
+  - A generic configuration profile to be used with [Podman](https://podman.io/)
+- `shifter`
+  - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/)
+- `charliecloud`
+  - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/)
+- `conda`
+  - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud.
+
+### `-resume`
+
+Specify this when restarting a pipeline. Nextflow will use cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. For input to be considered the same, not only the names must be identical but the files' contents as well. For more info about this parameter, see [this blog post](https://www.nextflow.io/blog/2019/demystifying-nextflow-resume.html).
+
+You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names.
+
+### `-c`
+
+Specify the path to a specific config file (this is a core Nextflow command). See the [nf-core website documentation](https://nf-co.re/usage/configuration) for more information.
+
+## Custom configuration
+
+### Resource requests
+
+Whilst the default requirements set within the pipeline will hopefully work for most people and with most input data, you may find that you want to customise the compute resources that the pipeline requests. Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with any of the error codes specified [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L18) it will automatically be resubmitted with higher requests (2 x original, then 3 x original). If it still fails after the third attempt then the pipeline execution is stopped.
+
+For example, if the nf-core/rnaseq pipeline is failing after multiple re-submissions of the `STAR_ALIGN` process due to an exit code of `137` this would indicate that there is an out of memory issue:
+
+```console
+[62/149eb0] NOTE: Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) -- Execution is retried (1)
+Error executing process > 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)'
+
+Caused by:
+    Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137)
+
+Command executed:
+    STAR \
+        --genomeDir star \
+        --readFilesIn WT_REP1_trimmed.fq.gz  \
+        --runThreadN 2 \
+        --outFileNamePrefix WT_REP1. \
+        <TRUNCATED>
+
+Command exit status:
+    137
+
+Command output:
+    (empty)
+
+Command error:
+    .command.sh: line 9:  30 Killed    STAR --genomeDir star --readFilesIn WT_REP1_trimmed.fq.gz --runThreadN 2 --outFileNamePrefix WT_REP1. <TRUNCATED>
+Work dir:
+    /home/pipelinetest/work/9d/172ca5881234073e8d76f2a19c88fb
+
+Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run`
+```
+
+#### For beginners
+
+A first step to bypass this error, you could try to increase the amount of CPUs, memory, and time for the whole pipeline. Therefor you can try to increase the resource for the parameters `--max_cpus`, `--max_memory`, and `--max_time`. Based on the error above, you have to increase the amount of memory. Therefore you can go to the [parameter documentation of rnaseq](https://nf-co.re/rnaseq/3.9/parameters) and scroll down to the `show hidden parameter` button to get the default value for `--max_memory`. In this case 128GB, you than can try to run your pipeline again with `--max_memory 200GB -resume` to skip all process, that were already calculated. If you can not increase the resource of the complete pipeline, you can try to adapt the resource for a single process as mentioned below.
+
+#### Advanced option on process level
+
+To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN).
+We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so, based on the search results, the file we want is `modules/nf-core/star/align/main.nf`.
+If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9).
+The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements.
+The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB.
+Providing you haven't set any other standard nf-core parameters to **cap** the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB.
+The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections.
+
+```nextflow
+process {
+    withName: 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN' {
+        memory = 100.GB
+    }
+}
+```
+
+> **NB:** We specify the full process name i.e. `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN` in the config file because this takes priority over the short name (`STAR_ALIGN`) and allows existing configuration using the full process name to be correctly overridden.
+>
+> If you get a warning suggesting that the process selector isn't recognised check that the process name has been specified correctly.
+
+### Updating containers (advanced users)
+
+The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. If for some reason you need to use a different version of a particular tool with the pipeline then you just need to identify the `process` name and override the Nextflow `container` definition for that process using the `withName` declaration. For example, in the [nf-core/viralrecon](https://nf-co.re/viralrecon) pipeline a tool called [Pangolin](https://github.com/cov-lineages/pangolin) has been used during the COVID-19 pandemic to assign lineages to SARS-CoV-2 genome sequenced samples. Given that the lineage assignments change quite frequently it doesn't make sense to re-release the nf-core/viralrecon everytime a new version of Pangolin has been released. However, you can override the default container used by the pipeline by creating a custom config file and passing it as a command-line argument via `-c custom.config`.
+
+1. Check the default version used by the pipeline in the module file for [Pangolin](https://github.com/nf-core/viralrecon/blob/a85d5969f9025409e3618d6c280ef15ce417df65/modules/nf-core/software/pangolin/main.nf#L14-L19)
+2. Find the latest version of the Biocontainer available on [Quay.io](https://quay.io/repository/biocontainers/pangolin?tag=latest&tab=tags)
+3. Create the custom config accordingly:
+
+   - For Docker:
+
+     ```nextflow
+     process {
+         withName: PANGOLIN {
+             container = 'quay.io/biocontainers/pangolin:3.0.5--pyhdfd78af_0'
+         }
+     }
+     ```
+
+   - For Singularity:
+
+     ```nextflow
+     process {
+         withName: PANGOLIN {
+             container = 'https://depot.galaxyproject.org/singularity/pangolin:3.0.5--pyhdfd78af_0'
+         }
+     }
+     ```
+
+   - For Conda:
+
+     ```nextflow
+     process {
+         withName: PANGOLIN {
+             conda = 'bioconda::pangolin=3.0.5'
+         }
+     }
+     ```
+
+> **NB:** If you wish to periodically update individual tool-specific results (e.g. Pangolin) generated by the pipeline then you must ensure to keep the `work/` directory otherwise the `-resume` ability of the pipeline will be compromised and it will restart from scratch.
+
+### nf-core/configs
+
+In most cases, you will only need to create a custom config as a one-off but if you and others within your organisation are likely to be running nf-core pipelines regularly and need to use the same settings regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter. You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile.
+
+See the main [Nextflow documentation](https://www.nextflow.io/docs/latest/config.html) for more information about creating your own configuration files.
+
+If you have any questions or issues please send us a message on [Slack](https://nf-co.re/join/slack) on the [`#configs` channel](https://nfcore.slack.com/channels/configs).
+
+## Azure Resource Requests
+
+To be used with the `azurebatch` profile by specifying the `-profile azurebatch`.
+We recommend providing a compute `params.vm_type` of `Standard_D16_v3` VMs by default but these options can be changed if required.
+
+Note that the choice of VM size depends on your quota and the overall workload during the analysis.
+For a thorough list, please refer the [Azure Sizes for virtual machines in Azure](https://docs.microsoft.com/en-us/azure/virtual-machines/sizes).
+
+## Running in the background
+
+Nextflow handles job submissions and supervises the running jobs. The Nextflow process must run until the pipeline is finished.
+
+The Nextflow `-bg` flag launches Nextflow in the background, detached from your terminal so that the workflow does not stop if you log out of your session. The logs are saved to a file.
+
+Alternatively, you can use `screen` / `tmux` or similar tool to create a detached session which you can log back into at a later time.
+Some HPC setups also allow you to run nextflow within a cluster job submitted your job scheduler (from where it submits more jobs).
+
+## Nextflow memory requirements
+
+In some cases, the Nextflow Java virtual machines can start to request a large amount of memory.
+We recommend adding the following line to your environment to limit this (typically in `~/.bashrc` or `~./bash_profile`):
+
+```bash
+NXF_OPTS='-Xms1g -Xmx4g'
+```
diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy
new file mode 100755
index 00000000..33cd4f6e
--- /dev/null
+++ b/lib/NfcoreSchema.groovy
@@ -0,0 +1,528 @@
+//
+// This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template.
+//
+
+import org.everit.json.schema.Schema
+import org.everit.json.schema.loader.SchemaLoader
+import org.everit.json.schema.ValidationException
+import org.json.JSONObject
+import org.json.JSONTokener
+import org.json.JSONArray
+import groovy.json.JsonSlurper
+import groovy.json.JsonBuilder
+
+class NfcoreSchema {
+
+    //
+    // Resolve Schema path relative to main workflow directory
+    //
+    public static String getSchemaPath(workflow, schema_filename='nextflow_schema.json') {
+        return "${workflow.projectDir}/${schema_filename}"
+    }
+
+    //
+    // Function to loop over all parameters defined in schema and check
+    // whether the given parameters adhere to the specifications
+    //
+    /* groovylint-disable-next-line UnusedPrivateMethodParameter */
+    public static void validateParameters(workflow, params, log, schema_filename='nextflow_schema.json') {
+        def has_error = false
+        //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~//
+        // Check for nextflow core params and unexpected params
+        def json = new File(getSchemaPath(workflow, schema_filename=schema_filename)).text
+        def Map schemaParams = (Map) new JsonSlurper().parseText(json).get('definitions')
+        def nf_params = [
+            // Options for base `nextflow` command
+            'bg',
+            'c',
+            'C',
+            'config',
+            'd',
+            'D',
+            'dockerize',
+            'h',
+            'log',
+            'q',
+            'quiet',
+            'syslog',
+            'v',
+
+            // Options for `nextflow run` command
+            'ansi',
+            'ansi-log',
+            'bg',
+            'bucket-dir',
+            'c',
+            'cache',
+            'config',
+            'dsl2',
+            'dump-channels',
+            'dump-hashes',
+            'E',
+            'entry',
+            'latest',
+            'lib',
+            'main-script',
+            'N',
+            'name',
+            'offline',
+            'params-file',
+            'pi',
+            'plugins',
+            'poll-interval',
+            'pool-size',
+            'profile',
+            'ps',
+            'qs',
+            'queue-size',
+            'r',
+            'resume',
+            'revision',
+            'stdin',
+            'stub',
+            'stub-run',
+            'test',
+            'w',
+            'with-charliecloud',
+            'with-conda',
+            'with-dag',
+            'with-docker',
+            'with-mpi',
+            'with-notification',
+            'with-podman',
+            'with-report',
+            'with-singularity',
+            'with-timeline',
+            'with-tower',
+            'with-trace',
+            'with-weblog',
+            'without-docker',
+            'without-podman',
+            'work-dir'
+        ]
+        def unexpectedParams = []
+
+        // Collect expected parameters from the schema
+        def expectedParams = []
+        def enums = [:]
+        for (group in schemaParams) {
+            for (p in group.value['properties']) {
+                expectedParams.push(p.key)
+                if (group.value['properties'][p.key].containsKey('enum')) {
+                    enums[p.key] = group.value['properties'][p.key]['enum']
+                }
+            }
+        }
+
+        for (specifiedParam in params.keySet()) {
+            // nextflow params
+            if (nf_params.contains(specifiedParam)) {
+                log.error "ERROR: You used a core Nextflow option with two hyphens: '--${specifiedParam}'. Please resubmit with '-${specifiedParam}'"
+                has_error = true
+            }
+            // unexpected params
+            def params_ignore = params.schema_ignore_params.split(',') + 'schema_ignore_params'
+            def expectedParamsLowerCase = expectedParams.collect{ it.replace("-", "").toLowerCase() }
+            def specifiedParamLowerCase = specifiedParam.replace("-", "").toLowerCase()
+            def isCamelCaseBug = (specifiedParam.contains("-") && !expectedParams.contains(specifiedParam) && expectedParamsLowerCase.contains(specifiedParamLowerCase))
+            if (!expectedParams.contains(specifiedParam) && !params_ignore.contains(specifiedParam) && !isCamelCaseBug) {
+                // Temporarily remove camelCase/camel-case params #1035
+                def unexpectedParamsLowerCase = unexpectedParams.collect{ it.replace("-", "").toLowerCase()}
+                if (!unexpectedParamsLowerCase.contains(specifiedParamLowerCase)){
+                    unexpectedParams.push(specifiedParam)
+                }
+            }
+        }
+
+        //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~//
+        // Validate parameters against the schema
+        InputStream input_stream = new File(getSchemaPath(workflow, schema_filename=schema_filename)).newInputStream()
+        JSONObject raw_schema = new JSONObject(new JSONTokener(input_stream))
+
+        // Remove anything that's in params.schema_ignore_params
+        raw_schema = removeIgnoredParams(raw_schema, params)
+
+        Schema schema = SchemaLoader.load(raw_schema)
+
+        // Clean the parameters
+        def cleanedParams = cleanParameters(params)
+
+        // Convert to JSONObject
+        def jsonParams = new JsonBuilder(cleanedParams)
+        JSONObject params_json = new JSONObject(jsonParams.toString())
+
+        // Validate
+        try {
+            schema.validate(params_json)
+        } catch (ValidationException e) {
+            println ''
+            log.error 'ERROR: Validation of pipeline parameters failed!'
+            JSONObject exceptionJSON = e.toJSON()
+            printExceptions(exceptionJSON, params_json, log, enums)
+            println ''
+            has_error = true
+        }
+
+        // Check for unexpected parameters
+        if (unexpectedParams.size() > 0) {
+            Map colors = NfcoreTemplate.logColours(params.monochrome_logs)
+            println ''
+            def warn_msg = 'Found unexpected parameters:'
+            for (unexpectedParam in unexpectedParams) {
+                warn_msg = warn_msg + "\n* --${unexpectedParam}: ${params[unexpectedParam].toString()}"
+            }
+            log.warn warn_msg
+            log.info "- ${colors.dim}Ignore this warning: params.schema_ignore_params = \"${unexpectedParams.join(',')}\" ${colors.reset}"
+            println ''
+        }
+
+        if (has_error) {
+            System.exit(1)
+        }
+    }
+
+    //
+    // Beautify parameters for --help
+    //
+    public static String paramsHelp(workflow, params, command, schema_filename='nextflow_schema.json') {
+        Map colors = NfcoreTemplate.logColours(params.monochrome_logs)
+        Integer num_hidden = 0
+        String output  = ''
+        output        += 'Typical pipeline command:\n\n'
+        output        += "  ${colors.cyan}${command}${colors.reset}\n\n"
+        Map params_map = paramsLoad(getSchemaPath(workflow, schema_filename=schema_filename))
+        Integer max_chars  = paramsMaxChars(params_map) + 1
+        Integer desc_indent = max_chars + 14
+        Integer dec_linewidth = 160 - desc_indent
+        for (group in params_map.keySet()) {
+            Integer num_params = 0
+            String group_output = colors.underlined + colors.bold + group + colors.reset + '\n'
+            def group_params = params_map.get(group)  // This gets the parameters of that particular group
+            for (param in group_params.keySet()) {
+                if (group_params.get(param).hidden && !params.show_hidden_params) {
+                    num_hidden += 1
+                    continue;
+                }
+                def type = '[' + group_params.get(param).type + ']'
+                def description = group_params.get(param).description
+                def defaultValue = group_params.get(param).default != null ? " [default: " + group_params.get(param).default.toString() + "]" : ''
+                def description_default = description + colors.dim + defaultValue + colors.reset
+                // Wrap long description texts
+                // Loosely based on https://dzone.com/articles/groovy-plain-text-word-wrap
+                if (description_default.length() > dec_linewidth){
+                    List olines = []
+                    String oline = "" // " " * indent
+                    description_default.split(" ").each() { wrd ->
+                        if ((oline.size() + wrd.size()) <= dec_linewidth) {
+                            oline += wrd + " "
+                        } else {
+                            olines += oline
+                            oline = wrd + " "
+                        }
+                    }
+                    olines += oline
+                    description_default = olines.join("\n" + " " * desc_indent)
+                }
+                group_output += "  --" +  param.padRight(max_chars) + colors.dim + type.padRight(10) + colors.reset + description_default + '\n'
+                num_params += 1
+            }
+            group_output += '\n'
+            if (num_params > 0){
+                output += group_output
+            }
+        }
+        if (num_hidden > 0){
+            output += colors.dim + "!! Hiding $num_hidden params, use --show_hidden_params to show them !!\n" + colors.reset
+        }
+        output += NfcoreTemplate.dashedLine(params.monochrome_logs)
+        return output
+    }
+
+    //
+    // Groovy Map summarising parameters/workflow options used by the pipeline
+    //
+    public static LinkedHashMap paramsSummaryMap(workflow, params, schema_filename='nextflow_schema.json') {
+        // Get a selection of core Nextflow workflow options
+        def Map workflow_summary = [:]
+        if (workflow.revision) {
+            workflow_summary['revision'] = workflow.revision
+        }
+        workflow_summary['runName']      = workflow.runName
+        if (workflow.containerEngine) {
+            workflow_summary['containerEngine'] = workflow.containerEngine
+        }
+        if (workflow.container) {
+            workflow_summary['container'] = workflow.container
+        }
+        workflow_summary['launchDir']    = workflow.launchDir
+        workflow_summary['workDir']      = workflow.workDir
+        workflow_summary['projectDir']   = workflow.projectDir
+        workflow_summary['userName']     = workflow.userName
+        workflow_summary['profile']      = workflow.profile
+        workflow_summary['configFiles']  = workflow.configFiles.join(', ')
+
+        // Get pipeline parameters defined in JSON Schema
+        def Map params_summary = [:]
+        def params_map = paramsLoad(getSchemaPath(workflow, schema_filename=schema_filename))
+        for (group in params_map.keySet()) {
+            def sub_params = new LinkedHashMap()
+            def group_params = params_map.get(group)  // This gets the parameters of that particular group
+            for (param in group_params.keySet()) {
+                if (params.containsKey(param)) {
+                    def params_value = params.get(param)
+                    def schema_value = group_params.get(param).default
+                    def param_type   = group_params.get(param).type
+                    if (schema_value != null) {
+                        if (param_type == 'string') {
+                            if (schema_value.contains('$projectDir') || schema_value.contains('${projectDir}')) {
+                                def sub_string = schema_value.replace('\$projectDir', '')
+                                sub_string     = sub_string.replace('\${projectDir}', '')
+                                if (params_value.contains(sub_string)) {
+                                    schema_value = params_value
+                                }
+                            }
+                            if (schema_value.contains('$params.outdir') || schema_value.contains('${params.outdir}')) {
+                                def sub_string = schema_value.replace('\$params.outdir', '')
+                                sub_string     = sub_string.replace('\${params.outdir}', '')
+                                if ("${params.outdir}${sub_string}" == params_value) {
+                                    schema_value = params_value
+                                }
+                            }
+                        }
+                    }
+
+                    // We have a default in the schema, and this isn't it
+                    if (schema_value != null && params_value != schema_value) {
+                        sub_params.put(param, params_value)
+                    }
+                    // No default in the schema, and this isn't empty
+                    else if (schema_value == null && params_value != "" && params_value != null && params_value != false) {
+                        sub_params.put(param, params_value)
+                    }
+                }
+            }
+            params_summary.put(group, sub_params)
+        }
+        return [ 'Core Nextflow options' : workflow_summary ] << params_summary
+    }
+
+    //
+    // Beautify parameters for summary and return as string
+    //
+    public static String paramsSummaryLog(workflow, params) {
+        Map colors = NfcoreTemplate.logColours(params.monochrome_logs)
+        String output  = ''
+        def params_map = paramsSummaryMap(workflow, params)
+        def max_chars  = paramsMaxChars(params_map)
+        for (group in params_map.keySet()) {
+            def group_params = params_map.get(group)  // This gets the parameters of that particular group
+            if (group_params) {
+                output += colors.bold + group + colors.reset + '\n'
+                for (param in group_params.keySet()) {
+                    output += "  " + colors.blue + param.padRight(max_chars) + ": " + colors.green +  group_params.get(param) + colors.reset + '\n'
+                }
+                output += '\n'
+            }
+        }
+        output += "!! Only displaying parameters that differ from the pipeline defaults !!\n"
+        output += NfcoreTemplate.dashedLine(params.monochrome_logs)
+        return output
+    }
+
+    //
+    // Loop over nested exceptions and print the causingException
+    //
+    private static void printExceptions(ex_json, params_json, log, enums, limit=5) {
+        def causingExceptions = ex_json['causingExceptions']
+        if (causingExceptions.length() == 0) {
+            def m = ex_json['message'] =~ /required key \[([^\]]+)\] not found/
+            // Missing required param
+            if (m.matches()) {
+                log.error "* Missing required parameter: --${m[0][1]}"
+            }
+            // Other base-level error
+            else if (ex_json['pointerToViolation'] == '#') {
+                log.error "* ${ex_json['message']}"
+            }
+            // Error with specific param
+            else {
+                def param = ex_json['pointerToViolation'] - ~/^#\//
+                def param_val = params_json[param].toString()
+                if (enums.containsKey(param)) {
+                    def error_msg = "* --${param}: '${param_val}' is not a valid choice (Available choices"
+                    if (enums[param].size() > limit) {
+                        log.error "${error_msg} (${limit} of ${enums[param].size()}): ${enums[param][0..limit-1].join(', ')}, ... )"
+                    } else {
+                        log.error "${error_msg}: ${enums[param].join(', ')})"
+                    }
+                } else {
+                    log.error "* --${param}: ${ex_json['message']} (${param_val})"
+                }
+            }
+        }
+        for (ex in causingExceptions) {
+            printExceptions(ex, params_json, log, enums)
+        }
+    }
+
+    //
+    // Remove an element from a JSONArray
+    //
+    private static JSONArray removeElement(json_array, element) {
+        def list = []
+        int len = json_array.length()
+        for (int i=0;i<len;i++){
+            list.add(json_array.get(i).toString())
+        }
+        list.remove(element)
+        JSONArray jsArray = new JSONArray(list)
+        return jsArray
+    }
+
+    //
+    // Remove ignored parameters
+    //
+    private static JSONObject removeIgnoredParams(raw_schema, params) {
+        // Remove anything that's in params.schema_ignore_params
+        params.schema_ignore_params.split(',').each{ ignore_param ->
+            if(raw_schema.keySet().contains('definitions')){
+                raw_schema.definitions.each { definition ->
+                    for (key in definition.keySet()){
+                        if (definition[key].get("properties").keySet().contains(ignore_param)){
+                            // Remove the param to ignore
+                            definition[key].get("properties").remove(ignore_param)
+                            // If the param was required, change this
+                            if (definition[key].has("required")) {
+                                def cleaned_required = removeElement(definition[key].required, ignore_param)
+                                definition[key].put("required", cleaned_required)
+                            }
+                        }
+                    }
+                }
+            }
+            if(raw_schema.keySet().contains('properties') && raw_schema.get('properties').keySet().contains(ignore_param)) {
+                raw_schema.get("properties").remove(ignore_param)
+            }
+            if(raw_schema.keySet().contains('required') && raw_schema.required.contains(ignore_param)) {
+                def cleaned_required = removeElement(raw_schema.required, ignore_param)
+                raw_schema.put("required", cleaned_required)
+            }
+        }
+        return raw_schema
+    }
+
+    //
+    // Clean and check parameters relative to Nextflow native classes
+    //
+    private static Map cleanParameters(params) {
+        def new_params = params.getClass().newInstance(params)
+        for (p in params) {
+            // remove anything evaluating to false
+            if (!p['value']) {
+                new_params.remove(p.key)
+            }
+            // Cast MemoryUnit to String
+            if (p['value'].getClass() == nextflow.util.MemoryUnit) {
+                new_params.replace(p.key, p['value'].toString())
+            }
+            // Cast Duration to String
+            if (p['value'].getClass() == nextflow.util.Duration) {
+                new_params.replace(p.key, p['value'].toString().replaceFirst(/d(?!\S)/, "day"))
+            }
+            // Cast LinkedHashMap to String
+            if (p['value'].getClass() == LinkedHashMap) {
+                new_params.replace(p.key, p['value'].toString())
+            }
+        }
+        return new_params
+    }
+
+    //
+    // This function tries to read a JSON params file
+    //
+    private static LinkedHashMap paramsLoad(String json_schema) {
+        def params_map = new LinkedHashMap()
+        try {
+            params_map = paramsRead(json_schema)
+        } catch (Exception e) {
+            println "Could not read parameters settings from JSON. $e"
+            params_map = new LinkedHashMap()
+        }
+        return params_map
+    }
+
+    //
+    // Method to actually read in JSON file using Groovy.
+    // Group (as Key), values are all parameters
+    //    - Parameter1 as Key, Description as Value
+    //    - Parameter2 as Key, Description as Value
+    //    ....
+    // Group
+    //    -
+    private static LinkedHashMap paramsRead(String json_schema) throws Exception {
+        def json = new File(json_schema).text
+        def Map schema_definitions = (Map) new JsonSlurper().parseText(json).get('definitions')
+        def Map schema_properties = (Map) new JsonSlurper().parseText(json).get('properties')
+        /* Tree looks like this in nf-core schema
+        * definitions <- this is what the first get('definitions') gets us
+                group 1
+                    title
+                    description
+                        properties
+                        parameter 1
+                            type
+                            description
+                        parameter 2
+                            type
+                            description
+                group 2
+                    title
+                    description
+                        properties
+                        parameter 1
+                            type
+                            description
+        * properties <- parameters can also be ungrouped, outside of definitions
+                parameter 1
+                    type
+                    description
+        */
+
+        // Grouped params
+        def params_map = new LinkedHashMap()
+        schema_definitions.each { key, val ->
+            def Map group = schema_definitions."$key".properties // Gets the property object of the group
+            def title = schema_definitions."$key".title
+            def sub_params = new LinkedHashMap()
+            group.each { innerkey, value ->
+                sub_params.put(innerkey, value)
+            }
+            params_map.put(title, sub_params)
+        }
+
+        // Ungrouped params
+        def ungrouped_params = new LinkedHashMap()
+        schema_properties.each { innerkey, value ->
+            ungrouped_params.put(innerkey, value)
+        }
+        params_map.put("Other parameters", ungrouped_params)
+
+        return params_map
+    }
+
+    //
+    // Get maximum number of characters across all parameter names
+    //
+    private static Integer paramsMaxChars(params_map) {
+        Integer max_chars = 0
+        for (group in params_map.keySet()) {
+            def group_params = params_map.get(group)  // This gets the parameters of that particular group
+            for (param in group_params.keySet()) {
+                if (param.size() > max_chars) {
+                    max_chars = param.size()
+                }
+            }
+        }
+        return max_chars
+    }
+}
diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
new file mode 100755
index 00000000..25a0a74a
--- /dev/null
+++ b/lib/NfcoreTemplate.groovy
@@ -0,0 +1,336 @@
+//
+// This file holds several functions used within the nf-core pipeline template.
+//
+
+import org.yaml.snakeyaml.Yaml
+
+class NfcoreTemplate {
+
+    //
+    // Check AWS Batch related parameters have been specified correctly
+    //
+    public static void awsBatch(workflow, params) {
+        if (workflow.profile.contains('awsbatch')) {
+            // Check params.awsqueue and params.awsregion have been set if running on AWSBatch
+            assert (params.awsqueue && params.awsregion) : "Specify correct --awsqueue and --awsregion parameters on AWSBatch!"
+            // Check outdir paths to be S3 buckets if running on AWSBatch
+            assert params.outdir.startsWith('s3:')       : "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!"
+        }
+    }
+
+    //
+    //  Warn if a -profile or Nextflow config has not been provided to run the pipeline
+    //
+    public static void checkConfigProvided(workflow, log) {
+        if (workflow.profile == 'standard' && workflow.configFiles.size() <= 1) {
+            log.warn "[$workflow.manifest.name] You are attempting to run the pipeline without any custom configuration!\n\n" +
+                    "This will be dependent on your local compute environment but can be achieved via one or more of the following:\n" +
+                    "   (1) Using an existing pipeline profile e.g. `-profile docker` or `-profile singularity`\n" +
+                    "   (2) Using an existing nf-core/configs for your Institution e.g. `-profile crick` or `-profile uppmax`\n" +
+                    "   (3) Using your own local custom config e.g. `-c /path/to/your/custom.config`\n\n" +
+                    "Please refer to the quick start section and usage docs for the pipeline.\n "
+        }
+    }
+
+    //
+    // Generate version string
+    //
+    public static String version(workflow) {
+        String version_string = ""
+
+        if (workflow.manifest.version) {
+            def prefix_v = workflow.manifest.version[0] != 'v' ? 'v' : ''
+            version_string += "${prefix_v}${workflow.manifest.version}"
+        }
+
+        if (workflow.commitId) {
+            def git_shortsha = workflow.commitId.substring(0, 7)
+            version_string += "-g${git_shortsha}"
+        }
+
+        return version_string
+    }
+
+    //
+    // Construct and send completion email
+    //
+    public static void email(workflow, params, summary_params, projectDir, log, multiqc_report=[]) {
+
+        // Set up the e-mail variables
+        def subject = "[$workflow.manifest.name] Successful: $workflow.runName"
+        if (!workflow.success) {
+            subject = "[$workflow.manifest.name] FAILED: $workflow.runName"
+        }
+
+        def summary = [:]
+        for (group in summary_params.keySet()) {
+            summary << summary_params[group]
+        }
+
+        def misc_fields = [:]
+        misc_fields['Date Started']              = workflow.start
+        misc_fields['Date Completed']            = workflow.complete
+        misc_fields['Pipeline script file path'] = workflow.scriptFile
+        misc_fields['Pipeline script hash ID']   = workflow.scriptId
+        if (workflow.repository) misc_fields['Pipeline repository Git URL']    = workflow.repository
+        if (workflow.commitId)   misc_fields['Pipeline repository Git Commit'] = workflow.commitId
+        if (workflow.revision)   misc_fields['Pipeline Git branch/tag']        = workflow.revision
+        misc_fields['Nextflow Version']           = workflow.nextflow.version
+        misc_fields['Nextflow Build']             = workflow.nextflow.build
+        misc_fields['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp
+
+        def email_fields = [:]
+        email_fields['version']      = NfcoreTemplate.version(workflow)
+        email_fields['runName']      = workflow.runName
+        email_fields['success']      = workflow.success
+        email_fields['dateComplete'] = workflow.complete
+        email_fields['duration']     = workflow.duration
+        email_fields['exitStatus']   = workflow.exitStatus
+        email_fields['errorMessage'] = (workflow.errorMessage ?: 'None')
+        email_fields['errorReport']  = (workflow.errorReport ?: 'None')
+        email_fields['commandLine']  = workflow.commandLine
+        email_fields['projectDir']   = workflow.projectDir
+        email_fields['summary']      = summary << misc_fields
+
+        // On success try attach the multiqc report
+        def mqc_report = null
+        try {
+            if (workflow.success) {
+                mqc_report = multiqc_report.getVal()
+                if (mqc_report.getClass() == ArrayList && mqc_report.size() >= 1) {
+                    if (mqc_report.size() > 1) {
+                        log.warn "[$workflow.manifest.name] Found multiple reports from process 'MULTIQC', will use only one"
+                    }
+                    mqc_report = mqc_report[0]
+                }
+            }
+        } catch (all) {
+            if (multiqc_report) {
+                log.warn "[$workflow.manifest.name] Could not attach MultiQC report to summary email"
+            }
+        }
+
+        // Check if we are only sending emails on failure
+        def email_address = params.email
+        if (!params.email && params.email_on_fail && !workflow.success) {
+            email_address = params.email_on_fail
+        }
+
+        // Render the TXT template
+        def engine       = new groovy.text.GStringTemplateEngine()
+        def tf           = new File("$projectDir/assets/email_template.txt")
+        def txt_template = engine.createTemplate(tf).make(email_fields)
+        def email_txt    = txt_template.toString()
+
+        // Render the HTML template
+        def hf            = new File("$projectDir/assets/email_template.html")
+        def html_template = engine.createTemplate(hf).make(email_fields)
+        def email_html    = html_template.toString()
+
+        // Render the sendmail template
+        def max_multiqc_email_size = params.max_multiqc_email_size as nextflow.util.MemoryUnit
+        def smail_fields           = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ]
+        def sf                     = new File("$projectDir/assets/sendmail_template.txt")
+        def sendmail_template      = engine.createTemplate(sf).make(smail_fields)
+        def sendmail_html          = sendmail_template.toString()
+
+        // Send the HTML e-mail
+        Map colors = logColours(params.monochrome_logs)
+        if (email_address) {
+            try {
+                if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') }
+                // Try to send HTML e-mail using sendmail
+                [ 'sendmail', '-t' ].execute() << sendmail_html
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (sendmail)-"
+            } catch (all) {
+                // Catch failures and try with plaintext
+                def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ]
+                if ( mqc_report.size() <= max_multiqc_email_size.toBytes() ) {
+                    mail_cmd += [ '-A', mqc_report ]
+                }
+                mail_cmd.execute() << email_html
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (mail)-"
+            }
+        }
+
+        // Write summary e-mail HTML to a file
+        def output_d = new File("${params.outdir}/pipeline_info/")
+        if (!output_d.exists()) {
+            output_d.mkdirs()
+        }
+        def output_hf = new File(output_d, "pipeline_report.html")
+        output_hf.withWriter { w -> w << email_html }
+        def output_tf = new File(output_d, "pipeline_report.txt")
+        output_tf.withWriter { w -> w << email_txt }
+    }
+
+    //
+    // Construct and send a notification to a web server as JSON
+    // e.g. Microsoft Teams and Slack
+    //
+    public static void IM_notification(workflow, params, summary_params, projectDir, log) {
+        def hook_url = params.hook_url
+
+        def summary = [:]
+        for (group in summary_params.keySet()) {
+            summary << summary_params[group]
+        }
+
+        def misc_fields = [:]
+        misc_fields['start']                                = workflow.start
+        misc_fields['complete']                             = workflow.complete
+        misc_fields['scriptfile']                           = workflow.scriptFile
+        misc_fields['scriptid']                             = workflow.scriptId
+        if (workflow.repository) misc_fields['repository']  = workflow.repository
+        if (workflow.commitId)   misc_fields['commitid']    = workflow.commitId
+        if (workflow.revision)   misc_fields['revision']    = workflow.revision
+        misc_fields['nxf_version']                          = workflow.nextflow.version
+        misc_fields['nxf_build']                            = workflow.nextflow.build
+        misc_fields['nxf_timestamp']                        = workflow.nextflow.timestamp
+
+        def msg_fields = [:]
+        msg_fields['version']      = NfcoreTemplate.version(workflow)
+        msg_fields['runName']      = workflow.runName
+        msg_fields['success']      = workflow.success
+        msg_fields['dateComplete'] = workflow.complete
+        msg_fields['duration']     = workflow.duration
+        msg_fields['exitStatus']   = workflow.exitStatus
+        msg_fields['errorMessage'] = (workflow.errorMessage ?: 'None')
+        msg_fields['errorReport']  = (workflow.errorReport ?: 'None')
+        msg_fields['commandLine']  = workflow.commandLine.replaceFirst(/ +--hook_url +[^ ]+/, "")
+        msg_fields['projectDir']   = workflow.projectDir
+        msg_fields['summary']      = summary << misc_fields
+
+        // Render the JSON template
+        def engine       = new groovy.text.GStringTemplateEngine()
+        // Different JSON depending on the service provider
+        // Defaults to "Adaptive Cards" (https://adaptivecards.io), except Slack which has its own format
+        def json_path     = hook_url.contains("hooks.slack.com") ? "slackreport.json" : "adaptivecard.json"
+        def hf            = new File("$projectDir/assets/${json_path}")
+        def json_template = engine.createTemplate(hf).make(msg_fields)
+        def json_message  = json_template.toString()
+
+        // POST
+        def post = new URL(hook_url).openConnection();
+        post.setRequestMethod("POST")
+        post.setDoOutput(true)
+        post.setRequestProperty("Content-Type", "application/json")
+        post.getOutputStream().write(json_message.getBytes("UTF-8"));
+        def postRC = post.getResponseCode();
+        if (! postRC.equals(200)) {
+            log.warn(post.getErrorStream().getText());
+        }
+    }
+
+    //
+    // Print pipeline summary on completion
+    //
+    public static void summary(workflow, params, log) {
+        Map colors = logColours(params.monochrome_logs)
+        if (workflow.success) {
+            if (workflow.stats.ignoredCount == 0) {
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Pipeline completed successfully${colors.reset}-"
+            } else {
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.yellow} Pipeline completed successfully, but with errored process(es) ${colors.reset}-"
+            }
+        } else {
+            log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed with errors${colors.reset}-"
+        }
+    }
+
+    //
+    // ANSII Colours used for terminal logging
+    //
+    public static Map logColours(Boolean monochrome_logs) {
+        Map colorcodes = [:]
+
+        // Reset / Meta
+        colorcodes['reset']      = monochrome_logs ? '' : "\033[0m"
+        colorcodes['bold']       = monochrome_logs ? '' : "\033[1m"
+        colorcodes['dim']        = monochrome_logs ? '' : "\033[2m"
+        colorcodes['underlined'] = monochrome_logs ? '' : "\033[4m"
+        colorcodes['blink']      = monochrome_logs ? '' : "\033[5m"
+        colorcodes['reverse']    = monochrome_logs ? '' : "\033[7m"
+        colorcodes['hidden']     = monochrome_logs ? '' : "\033[8m"
+
+        // Regular Colors
+        colorcodes['black']      = monochrome_logs ? '' : "\033[0;30m"
+        colorcodes['red']        = monochrome_logs ? '' : "\033[0;31m"
+        colorcodes['green']      = monochrome_logs ? '' : "\033[0;32m"
+        colorcodes['yellow']     = monochrome_logs ? '' : "\033[0;33m"
+        colorcodes['blue']       = monochrome_logs ? '' : "\033[0;34m"
+        colorcodes['purple']     = monochrome_logs ? '' : "\033[0;35m"
+        colorcodes['cyan']       = monochrome_logs ? '' : "\033[0;36m"
+        colorcodes['white']      = monochrome_logs ? '' : "\033[0;37m"
+
+        // Bold
+        colorcodes['bblack']     = monochrome_logs ? '' : "\033[1;30m"
+        colorcodes['bred']       = monochrome_logs ? '' : "\033[1;31m"
+        colorcodes['bgreen']     = monochrome_logs ? '' : "\033[1;32m"
+        colorcodes['byellow']    = monochrome_logs ? '' : "\033[1;33m"
+        colorcodes['bblue']      = monochrome_logs ? '' : "\033[1;34m"
+        colorcodes['bpurple']    = monochrome_logs ? '' : "\033[1;35m"
+        colorcodes['bcyan']      = monochrome_logs ? '' : "\033[1;36m"
+        colorcodes['bwhite']     = monochrome_logs ? '' : "\033[1;37m"
+
+        // Underline
+        colorcodes['ublack']     = monochrome_logs ? '' : "\033[4;30m"
+        colorcodes['ured']       = monochrome_logs ? '' : "\033[4;31m"
+        colorcodes['ugreen']     = monochrome_logs ? '' : "\033[4;32m"
+        colorcodes['uyellow']    = monochrome_logs ? '' : "\033[4;33m"
+        colorcodes['ublue']      = monochrome_logs ? '' : "\033[4;34m"
+        colorcodes['upurple']    = monochrome_logs ? '' : "\033[4;35m"
+        colorcodes['ucyan']      = monochrome_logs ? '' : "\033[4;36m"
+        colorcodes['uwhite']     = monochrome_logs ? '' : "\033[4;37m"
+
+        // High Intensity
+        colorcodes['iblack']     = monochrome_logs ? '' : "\033[0;90m"
+        colorcodes['ired']       = monochrome_logs ? '' : "\033[0;91m"
+        colorcodes['igreen']     = monochrome_logs ? '' : "\033[0;92m"
+        colorcodes['iyellow']    = monochrome_logs ? '' : "\033[0;93m"
+        colorcodes['iblue']      = monochrome_logs ? '' : "\033[0;94m"
+        colorcodes['ipurple']    = monochrome_logs ? '' : "\033[0;95m"
+        colorcodes['icyan']      = monochrome_logs ? '' : "\033[0;96m"
+        colorcodes['iwhite']     = monochrome_logs ? '' : "\033[0;97m"
+
+        // Bold High Intensity
+        colorcodes['biblack']    = monochrome_logs ? '' : "\033[1;90m"
+        colorcodes['bired']      = monochrome_logs ? '' : "\033[1;91m"
+        colorcodes['bigreen']    = monochrome_logs ? '' : "\033[1;92m"
+        colorcodes['biyellow']   = monochrome_logs ? '' : "\033[1;93m"
+        colorcodes['biblue']     = monochrome_logs ? '' : "\033[1;94m"
+        colorcodes['bipurple']   = monochrome_logs ? '' : "\033[1;95m"
+        colorcodes['bicyan']     = monochrome_logs ? '' : "\033[1;96m"
+        colorcodes['biwhite']    = monochrome_logs ? '' : "\033[1;97m"
+
+        return colorcodes
+    }
+
+    //
+    // Does what is says on the tin
+    //
+    public static String dashedLine(monochrome_logs) {
+        Map colors = logColours(monochrome_logs)
+        return "-${colors.dim}----------------------------------------------------${colors.reset}-"
+    }
+
+    //
+    // nf-core logo
+    //
+    public static String logo(workflow, monochrome_logs) {
+        Map colors = logColours(monochrome_logs)
+        String workflow_version = NfcoreTemplate.version(workflow)
+        String.format(
+            """\n
+            ${dashedLine(monochrome_logs)}
+                                                    ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset}
+            ${colors.blue}        ___     __   __   __   ___     ${colors.green}/,-._.--~\'${colors.reset}
+            ${colors.blue}  |\\ | |__  __ /  ` /  \\ |__) |__         ${colors.yellow}}  {${colors.reset}
+            ${colors.blue}  | \\| |       \\__, \\__/ |  \\ |___     ${colors.green}\\`-._,-`-,${colors.reset}
+                                                    ${colors.green}`._,._,\'${colors.reset}
+            ${colors.purple}  ${workflow.manifest.name} ${workflow_version}${colors.reset}
+            ${dashedLine(monochrome_logs)}
+            """.stripIndent()
+        )
+    }
+}
diff --git a/lib/Utils.groovy b/lib/Utils.groovy
new file mode 100644
index 00000000..8d030f4e
--- /dev/null
+++ b/lib/Utils.groovy
@@ -0,0 +1,47 @@
+//
+// This file holds several Groovy functions that could be useful for any Nextflow pipeline
+//
+
+import org.yaml.snakeyaml.Yaml
+
+class Utils {
+
+    //
+    // When running with -profile conda, warn if channels have not been set-up appropriately
+    //
+    public static void checkCondaChannels(log) {
+        Yaml parser = new Yaml()
+        def channels = []
+        try {
+            def config = parser.load("conda config --show channels".execute().text)
+            channels = config.channels
+        } catch(NullPointerException | IOException e) {
+            log.warn "Could not verify conda channel configuration."
+            return
+        }
+
+        // Check that all channels are present
+        // This channel list is ordered by required channel priority.
+        def required_channels_in_order = ['conda-forge', 'bioconda', 'defaults']
+        def channels_missing = ((required_channels_in_order as Set) - (channels as Set)) as Boolean
+
+        // Check that they are in the right order
+        def channel_priority_violation = false
+        def n = required_channels_in_order.size()
+        for (int i = 0; i < n - 1; i++) {
+            channel_priority_violation |= !(channels.indexOf(required_channels_in_order[i]) < channels.indexOf(required_channels_in_order[i+1]))
+        }
+
+        if (channels_missing | channel_priority_violation) {
+            log.warn "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" +
+                "  There is a problem with your Conda configuration!\n\n" +
+                "  You will need to set-up the conda-forge and bioconda channels correctly.\n" +
+                "  Please refer to https://bioconda.github.io/\n" +
+                "  The observed channel order is \n" +
+                "  ${channels}\n" +
+                "  but the following channel order is required:\n" +
+                "  ${required_channels_in_order}\n" +
+                "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~"
+        }
+    }
+}
diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy
new file mode 100755
index 00000000..6aeceecc
--- /dev/null
+++ b/lib/WorkflowMain.groovy
@@ -0,0 +1,99 @@
+//
+// This file holds several functions specific to the main.nf workflow in the nf-core/rnadeseq pipeline
+//
+
+class WorkflowMain {
+
+    //
+    // Citation string for pipeline
+    //
+    public static String citation(workflow) {
+        return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" +
+            // TODO nf-core: Add Zenodo DOI for pipeline after first release
+            //"* The pipeline\n" +
+            //"  https://doi.org/10.5281/zenodo.XXXXXXX\n\n" +
+            "* The nf-core framework\n" +
+            "  https://doi.org/10.1038/s41587-020-0439-x\n\n" +
+            "* Software dependencies\n" +
+            "  https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md"
+    }
+
+    //
+    // Generate help string
+    //
+    public static String help(workflow, params, log) {
+        def command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --genome GRCh37 -profile docker"
+        def help_string = ''
+        help_string += NfcoreTemplate.logo(workflow, params.monochrome_logs)
+        help_string += NfcoreSchema.paramsHelp(workflow, params, command)
+        help_string += '\n' + citation(workflow) + '\n'
+        help_string += NfcoreTemplate.dashedLine(params.monochrome_logs)
+        return help_string
+    }
+
+    //
+    // Generate parameter summary log string
+    //
+    public static String paramsSummaryLog(workflow, params, log) {
+        def summary_log = ''
+        summary_log += NfcoreTemplate.logo(workflow, params.monochrome_logs)
+        summary_log += NfcoreSchema.paramsSummaryLog(workflow, params)
+        summary_log += '\n' + citation(workflow) + '\n'
+        summary_log += NfcoreTemplate.dashedLine(params.monochrome_logs)
+        return summary_log
+    }
+
+    //
+    // Validate parameters and print summary to screen
+    //
+    public static void initialise(workflow, params, log) {
+        // Print help to screen if required
+        if (params.help) {
+            log.info help(workflow, params, log)
+            System.exit(0)
+        }
+
+        // Print workflow version and exit on --version
+        if (params.version) {
+            String workflow_version = NfcoreTemplate.version(workflow)
+            log.info "${workflow.manifest.name} ${workflow_version}"
+            System.exit(0)
+        }
+
+        // Print parameter summary log to screen
+        log.info paramsSummaryLog(workflow, params, log)
+
+        // Validate workflow parameters via the JSON schema
+        if (params.validate_params) {
+            NfcoreSchema.validateParameters(workflow, params, log)
+        }
+
+        // Check that a -profile or Nextflow config has been provided to run the pipeline
+        NfcoreTemplate.checkConfigProvided(workflow, log)
+
+        // Check that conda channels are set-up correctly
+        if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) {
+            Utils.checkCondaChannels(log)
+        }
+
+        // Check AWS batch settings
+        NfcoreTemplate.awsBatch(workflow, params)
+
+        // Check input has been provided
+        if (!params.input) {
+            log.error "Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'"
+            System.exit(1)
+        }
+    }
+    //
+    // Get attribute from genome config file e.g. fasta
+    //
+    public static Object getGenomeAttribute(params, attribute) {
+        if (params.genomes && params.genome && params.genomes.containsKey(params.genome)) {
+            if (params.genomes[ params.genome ].containsKey(attribute)) {
+                return params.genomes[ params.genome ][ attribute ]
+            }
+        }
+        return null
+    }
+}
diff --git a/lib/WorkflowRnadeseq.groovy b/lib/WorkflowRnadeseq.groovy
new file mode 100755
index 00000000..090a7663
--- /dev/null
+++ b/lib/WorkflowRnadeseq.groovy
@@ -0,0 +1,77 @@
+//
+// This file holds several functions specific to the workflow/rnadeseq.nf in the nf-core/rnadeseq pipeline
+//
+
+import groovy.text.SimpleTemplateEngine
+
+class WorkflowRnadeseq {
+
+    //
+    // Check and validate parameters
+    //
+    public static void initialise(params, log) {
+        genomeExistsError(params, log)
+
+
+        if (!params.fasta) {
+            log.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file."
+            System.exit(1)
+        }
+    }
+
+    //
+    // Get workflow summary for MultiQC
+    //
+    public static String paramsSummaryMultiqc(workflow, summary) {
+        String summary_section = ''
+        for (group in summary.keySet()) {
+            def group_params = summary.get(group)  // This gets the parameters of that particular group
+            if (group_params) {
+                summary_section += "    <p style=\"font-size:110%\"><b>$group</b></p>\n"
+                summary_section += "    <dl class=\"dl-horizontal\">\n"
+                for (param in group_params.keySet()) {
+                    summary_section += "        <dt>$param</dt><dd><samp>${group_params.get(param) ?: '<span style=\"color:#999999;\">N/A</a>'}</samp></dd>\n"
+                }
+                summary_section += "    </dl>\n"
+            }
+        }
+
+        String yaml_file_text  = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n"
+        yaml_file_text        += "description: ' - this information is collected when the pipeline is started.'\n"
+        yaml_file_text        += "section_name: '${workflow.manifest.name} Workflow Summary'\n"
+        yaml_file_text        += "section_href: 'https://github.com/${workflow.manifest.name}'\n"
+        yaml_file_text        += "plot_type: 'html'\n"
+        yaml_file_text        += "data: |\n"
+        yaml_file_text        += "${summary_section}"
+        return yaml_file_text
+    }
+
+    public static String methodsDescriptionText(run_workflow, mqc_methods_yaml) {
+        // Convert  to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file
+        def meta = [:]
+        meta.workflow = run_workflow.toMap()
+        meta["manifest_map"] = run_workflow.manifest.toMap()
+
+        meta["doi_text"] = meta.manifest_map.doi ? "(doi: <a href=\'https://doi.org/${meta.manifest_map.doi}\'>${meta.manifest_map.doi}</a>)" : ""
+        meta["nodoi_text"] = meta.manifest_map.doi ? "": "<li>If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used. </li>"
+
+        def methods_text = mqc_methods_yaml.text
+
+        def engine =  new SimpleTemplateEngine()
+        def description_html = engine.createTemplate(methods_text).make(meta)
+
+        return description_html
+    }//
+    // Exit pipeline if incorrect --genome key provided
+    //
+    private static void genomeExistsError(params, log) {
+        if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) {
+            log.error "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" +
+                "  Genome '${params.genome}' not found in any config files provided to the pipeline.\n" +
+                "  Currently, the available genome keys are:\n" +
+                "  ${params.genomes.keySet().join(", ")}\n" +
+                "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~"
+            System.exit(1)
+        }
+    }
+}
diff --git a/lib/nfcore_external_java_deps.jar b/lib/nfcore_external_java_deps.jar
new file mode 100644
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literal 0
HcmV?d00001

diff --git a/main.nf b/main.nf
new file mode 100644
index 00000000..96d9ce69
--- /dev/null
+++ b/main.nf
@@ -0,0 +1,64 @@
+#!/usr/bin/env nextflow
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Github : https://github.com/nf-core/rnadeseq
+
+    Website: https://nf-co.re/rnadeseq
+    Slack  : https://nfcore.slack.com/channels/rnadeseq
+----------------------------------------------------------------------------------------
+*/
+
+nextflow.enable.dsl = 2
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    GENOME PARAMETER VALUES
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+params.fasta = WorkflowMain.getGenomeAttribute(params, 'fasta')
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    VALIDATE & PRINT PARAMETER SUMMARY
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+WorkflowMain.initialise(workflow, params, log)
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    NAMED WORKFLOW FOR PIPELINE
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+include { RNADESEQ } from './workflows/rnadeseq'
+
+//
+// WORKFLOW: Run main nf-core/rnadeseq analysis pipeline
+//
+workflow NFCORE_RNADESEQ {
+    RNADESEQ ()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    RUN ALL WORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// WORKFLOW: Execute a single named workflow for the pipeline
+// See: https://github.com/nf-core/rnaseq/issues/619
+//
+workflow {
+    NFCORE_RNADESEQ ()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    THE END
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
diff --git a/modules.json b/modules.json
new file mode 100644
index 00000000..0d60c94b
--- /dev/null
+++ b/modules.json
@@ -0,0 +1,27 @@
+{
+    "name": "nf-core/rnadeseq",
+    "homePage": "https://github.com/nf-core/rnadeseq",
+    "repos": {
+        "https://github.com/nf-core/modules.git": {
+            "modules": {
+                "nf-core": {
+                    "custom/dumpsoftwareversions": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
+                    },
+                    "fastqc": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
+                    },
+                    "multiqc": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
+                    }
+                }
+            }
+        }
+    }
+}
diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf
new file mode 100644
index 00000000..a3c7232d
--- /dev/null
+++ b/modules/local/samplesheet_check.nf
@@ -0,0 +1,31 @@
+process SAMPLESHEET_CHECK {
+    tag "$samplesheet"
+    label 'process_single'
+
+    conda "conda-forge::python=3.8.3"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/python:3.8.3' :
+        'quay.io/biocontainers/python:3.8.3' }"
+
+    input:
+    path samplesheet
+
+    output:
+    path '*.csv'       , emit: csv
+    path "versions.yml", emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script: // This script is bundled with the pipeline, in nf-core/rnadeseq/bin/
+    """
+    check_samplesheet.py \\
+        $samplesheet \\
+        samplesheet.valid.csv
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        python: \$(python --version | sed 's/Python //g')
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/custom/dumpsoftwareversions/main.nf b/modules/nf-core/custom/dumpsoftwareversions/main.nf
new file mode 100644
index 00000000..3df21765
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/main.nf
@@ -0,0 +1,24 @@
+process CUSTOM_DUMPSOFTWAREVERSIONS {
+    label 'process_single'
+
+    // Requires `pyyaml` which does not have a dedicated container but is in the MultiQC container
+    conda "bioconda::multiqc=1.13"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/multiqc:1.13--pyhdfd78af_0' :
+        'quay.io/biocontainers/multiqc:1.13--pyhdfd78af_0' }"
+
+    input:
+    path versions
+
+    output:
+    path "software_versions.yml"    , emit: yml
+    path "software_versions_mqc.yml", emit: mqc_yml
+    path "versions.yml"             , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    template 'dumpsoftwareversions.py'
+}
diff --git a/modules/nf-core/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/custom/dumpsoftwareversions/meta.yml
new file mode 100644
index 00000000..60b546a0
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/meta.yml
@@ -0,0 +1,34 @@
+name: custom_dumpsoftwareversions
+description: Custom module used to dump software versions within the nf-core pipeline template
+keywords:
+  - custom
+  - version
+tools:
+  - custom:
+      description: Custom module used to dump software versions within the nf-core pipeline template
+      homepage: https://github.com/nf-core/tools
+      documentation: https://github.com/nf-core/tools
+      licence: ["MIT"]
+input:
+  - versions:
+      type: file
+      description: YML file containing software versions
+      pattern: "*.yml"
+
+output:
+  - yml:
+      type: file
+      description: Standard YML file containing software versions
+      pattern: "software_versions.yml"
+  - mqc_yml:
+      type: file
+      description: MultiQC custom content YML file containing software versions
+      pattern: "software_versions_mqc.yml"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+
+authors:
+  - "@drpatelh"
+  - "@grst"
diff --git a/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
new file mode 100755
index 00000000..e55b8d43
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
@@ -0,0 +1,102 @@
+#!/usr/bin/env python
+
+
+"""Provide functions to merge multiple versions.yml files."""
+
+
+import platform
+from textwrap import dedent
+
+import yaml
+
+
+def _make_versions_html(versions):
+    """Generate a tabular HTML output of all versions for MultiQC."""
+    html = [
+        dedent(
+            """\\
+            <style>
+            #nf-core-versions tbody:nth-child(even) {
+                background-color: #f2f2f2;
+            }
+            </style>
+            <table class="table" style="width:100%" id="nf-core-versions">
+                <thead>
+                    <tr>
+                        <th> Process Name </th>
+                        <th> Software </th>
+                        <th> Version  </th>
+                    </tr>
+                </thead>
+            """
+        )
+    ]
+    for process, tmp_versions in sorted(versions.items()):
+        html.append("<tbody>")
+        for i, (tool, version) in enumerate(sorted(tmp_versions.items())):
+            html.append(
+                dedent(
+                    f"""\\
+                    <tr>
+                        <td><samp>{process if (i == 0) else ''}</samp></td>
+                        <td><samp>{tool}</samp></td>
+                        <td><samp>{version}</samp></td>
+                    </tr>
+                    """
+                )
+            )
+        html.append("</tbody>")
+    html.append("</table>")
+    return "\\n".join(html)
+
+
+def main():
+    """Load all version files and generate merged output."""
+    versions_this_module = {}
+    versions_this_module["${task.process}"] = {
+        "python": platform.python_version(),
+        "yaml": yaml.__version__,
+    }
+
+    with open("$versions") as f:
+        versions_by_process = yaml.load(f, Loader=yaml.BaseLoader) | versions_this_module
+
+    # aggregate versions by the module name (derived from fully-qualified process name)
+    versions_by_module = {}
+    for process, process_versions in versions_by_process.items():
+        module = process.split(":")[-1]
+        try:
+            if versions_by_module[module] != process_versions:
+                raise AssertionError(
+                    "We assume that software versions are the same between all modules. "
+                    "If you see this error-message it means you discovered an edge-case "
+                    "and should open an issue in nf-core/tools. "
+                )
+        except KeyError:
+            versions_by_module[module] = process_versions
+
+    versions_by_module["Workflow"] = {
+        "Nextflow": "$workflow.nextflow.version",
+        "$workflow.manifest.name": "$workflow.manifest.version",
+    }
+
+    versions_mqc = {
+        "id": "software_versions",
+        "section_name": "${workflow.manifest.name} Software Versions",
+        "section_href": "https://github.com/${workflow.manifest.name}",
+        "plot_type": "html",
+        "description": "are collected at run time from the software output.",
+        "data": _make_versions_html(versions_by_module),
+    }
+
+    with open("software_versions.yml", "w") as f:
+        yaml.dump(versions_by_module, f, default_flow_style=False)
+    with open("software_versions_mqc.yml", "w") as f:
+        yaml.dump(versions_mqc, f, default_flow_style=False)
+
+    with open("versions.yml", "w") as f:
+        yaml.dump(versions_this_module, f, default_flow_style=False)
+
+
+if __name__ == "__main__":
+    main()
diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf
new file mode 100644
index 00000000..9ae58381
--- /dev/null
+++ b/modules/nf-core/fastqc/main.nf
@@ -0,0 +1,51 @@
+process FASTQC {
+    tag "$meta.id"
+    label 'process_medium'
+
+    conda "bioconda::fastqc=0.11.9"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0' :
+        'quay.io/biocontainers/fastqc:0.11.9--0' }"
+
+    input:
+    tuple val(meta), path(reads)
+
+    output:
+    tuple val(meta), path("*.html"), emit: html
+    tuple val(meta), path("*.zip") , emit: zip
+    path  "versions.yml"           , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    def prefix = task.ext.prefix ?: "${meta.id}"
+    // Make list of old name and new name pairs to use for renaming in the bash while loop
+    def old_new_pairs = reads instanceof Path || reads.size() == 1 ? [[ reads, "${prefix}.${reads.extension}" ]] : reads.withIndex().collect { entry, index -> [ entry, "${prefix}_${index + 1}.${entry.extension}" ] }
+    def rename_to = old_new_pairs*.join(' ').join(' ')
+    def renamed_files = old_new_pairs.collect{ old_name, new_name -> new_name }.join(' ')
+    """
+    printf "%s %s\\n" $rename_to | while read old_name new_name; do
+        [ -f "\${new_name}" ] || ln -s \$old_name \$new_name
+    done
+    fastqc $args --threads $task.cpus $renamed_files
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
+    END_VERSIONS
+    """
+
+    stub:
+    def prefix = task.ext.prefix ?: "${meta.id}"
+    """
+    touch ${prefix}.html
+    touch ${prefix}.zip
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/fastqc/meta.yml b/modules/nf-core/fastqc/meta.yml
new file mode 100644
index 00000000..4da5bb5a
--- /dev/null
+++ b/modules/nf-core/fastqc/meta.yml
@@ -0,0 +1,52 @@
+name: fastqc
+description: Run FastQC on sequenced reads
+keywords:
+  - quality control
+  - qc
+  - adapters
+  - fastq
+tools:
+  - fastqc:
+      description: |
+        FastQC gives general quality metrics about your reads.
+        It provides information about the quality score distribution
+        across your reads, the per base sequence content (%A/C/G/T).
+        You get information about adapter contamination and other
+        overrepresented sequences.
+      homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/
+      documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/
+      licence: ["GPL-2.0-only"]
+input:
+  - meta:
+      type: map
+      description: |
+        Groovy Map containing sample information
+        e.g. [ id:'test', single_end:false ]
+  - reads:
+      type: file
+      description: |
+        List of input FastQ files of size 1 and 2 for single-end and paired-end data,
+        respectively.
+output:
+  - meta:
+      type: map
+      description: |
+        Groovy Map containing sample information
+        e.g. [ id:'test', single_end:false ]
+  - html:
+      type: file
+      description: FastQC report
+      pattern: "*_{fastqc.html}"
+  - zip:
+      type: file
+      description: FastQC report archive
+      pattern: "*_{fastqc.zip}"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+authors:
+  - "@drpatelh"
+  - "@grst"
+  - "@ewels"
+  - "@FelixKrueger"
diff --git a/modules/nf-core/multiqc/main.nf b/modules/nf-core/multiqc/main.nf
new file mode 100644
index 00000000..68f66bea
--- /dev/null
+++ b/modules/nf-core/multiqc/main.nf
@@ -0,0 +1,53 @@
+process MULTIQC {
+    label 'process_single'
+
+    conda "bioconda::multiqc=1.13"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/multiqc:1.13--pyhdfd78af_0' :
+        'quay.io/biocontainers/multiqc:1.13--pyhdfd78af_0' }"
+
+    input:
+    path  multiqc_files, stageAs: "?/*"
+    path(multiqc_config)
+    path(extra_multiqc_config)
+    path(multiqc_logo)
+
+    output:
+    path "*multiqc_report.html", emit: report
+    path "*_data"              , emit: data
+    path "*_plots"             , optional:true, emit: plots
+    path "versions.yml"        , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    def config = multiqc_config ? "--config $multiqc_config" : ''
+    def extra_config = extra_multiqc_config ? "--config $extra_multiqc_config" : ''
+    """
+    multiqc \\
+        --force \\
+        $args \\
+        $config \\
+        $extra_config \\
+        .
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
+    END_VERSIONS
+    """
+
+    stub:
+    """
+    touch multiqc_data
+    touch multiqc_plots
+    touch multiqc_report.html
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/multiqc/meta.yml b/modules/nf-core/multiqc/meta.yml
new file mode 100644
index 00000000..ebc29b27
--- /dev/null
+++ b/modules/nf-core/multiqc/meta.yml
@@ -0,0 +1,55 @@
+name: MultiQC
+description: Aggregate results from bioinformatics analyses across many samples into a single report
+keywords:
+  - QC
+  - bioinformatics tools
+  - Beautiful stand-alone HTML report
+tools:
+  - multiqc:
+      description: |
+        MultiQC searches a given directory for analysis logs and compiles a HTML report.
+        It's a general use tool, perfect for summarising the output from numerous bioinformatics tools.
+      homepage: https://multiqc.info/
+      documentation: https://multiqc.info/docs/
+      licence: ["GPL-3.0-or-later"]
+
+input:
+  - multiqc_files:
+      type: file
+      description: |
+        List of reports / files recognised by MultiQC, for example the html and zip output of FastQC
+  - multiqc_config:
+      type: file
+      description: Optional config yml for MultiQC
+      pattern: "*.{yml,yaml}"
+  - extra_multiqc_config:
+      type: file
+      description: Second optional config yml for MultiQC. Will override common sections in multiqc_config.
+      pattern: "*.{yml,yaml}"
+  - multiqc_logo:
+      type: file
+      description: Optional logo file for MultiQC
+      pattern: "*.{png}"
+
+output:
+  - report:
+      type: file
+      description: MultiQC report file
+      pattern: "multiqc_report.html"
+  - data:
+      type: dir
+      description: MultiQC data dir
+      pattern: "multiqc_data"
+  - plots:
+      type: file
+      description: Plots created by MultiQC
+      pattern: "*_data"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+authors:
+  - "@abhi18av"
+  - "@bunop"
+  - "@drpatelh"
+  - "@jfy133"
diff --git a/nextflow.config b/nextflow.config
new file mode 100644
index 00000000..57918460
--- /dev/null
+++ b/nextflow.config
@@ -0,0 +1,234 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq Nextflow config file
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Default config options for all compute environments
+----------------------------------------------------------------------------------------
+*/
+
+// Global default params, used in configs
+params {
+
+    // TODO nf-core: Specify your pipeline's command line flags
+    // Input options
+    input                      = null
+
+
+    // References
+    genome                     = null
+    igenomes_base              = 's3://ngi-igenomes/igenomes'
+    igenomes_ignore            = false
+    // MultiQC options
+    multiqc_config             = null
+    multiqc_title              = null
+    multiqc_logo               = null
+    max_multiqc_email_size     = '25.MB'
+    multiqc_methods_description = null
+
+    // Boilerplate options
+    outdir                     = null
+    tracedir                   = "${params.outdir}/pipeline_info"
+    publish_dir_mode           = 'copy'
+    email                      = null
+    email_on_fail              = null
+    plaintext_email            = false
+    monochrome_logs            = false
+    hook_url                   = null
+    help                       = false
+    version                    = false
+    validate_params            = true
+    show_hidden_params         = false
+    schema_ignore_params       = 'genomes'
+
+
+    // Config options
+    custom_config_version      = 'master'
+    custom_config_base         = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}"
+    config_profile_description = null
+    config_profile_contact     = null
+    config_profile_url         = null
+    config_profile_name        = null
+
+
+    // Max resource options
+    // Defaults only, expecting to be overwritten
+    max_memory                 = '128.GB'
+    max_cpus                   = 16
+    max_time                   = '240.h'
+
+}
+
+// Load base.config by default for all pipelines
+includeConfig 'conf/base.config'
+
+// Load nf-core custom profiles from different Institutions
+try {
+    includeConfig "${params.custom_config_base}/nfcore_custom.config"
+} catch (Exception e) {
+    System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config")
+}
+
+// Load nf-core/rnadeseq custom profiles from different institutions.
+// Warning: Uncomment only if a pipeline-specific instititutional config already exists on nf-core/configs!
+// try {
+//   includeConfig "${params.custom_config_base}/pipeline/rnadeseq.config"
+// } catch (Exception e) {
+//   System.err.println("WARNING: Could not load nf-core/config/rnadeseq profiles: ${params.custom_config_base}/pipeline/rnadeseq.config")
+// }
+
+
+profiles {
+    debug { process.beforeScript = 'echo $HOSTNAME' }
+    conda {
+        conda.enabled          = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    mamba {
+        conda.enabled          = true
+        conda.useMamba         = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    docker {
+        docker.enabled         = true
+        docker.userEmulation   = true
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    arm {
+        docker.runOptions = '-u $(id -u):$(id -g) --platform=linux/amd64'
+    }
+    singularity {
+        singularity.enabled    = true
+        singularity.autoMounts = true
+        docker.enabled         = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    podman {
+        podman.enabled         = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    shifter {
+        shifter.enabled        = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        charliecloud.enabled   = false
+    }
+    charliecloud {
+        charliecloud.enabled   = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+    }
+    gitpod {
+        executor.name          = 'local'
+        executor.cpus          = 16
+        executor.memory        = 60.GB
+    }
+    test      { includeConfig 'conf/test.config'      }
+    test_full { includeConfig 'conf/test_full.config' }
+}
+
+
+// Load igenomes.config if required
+if (!params.igenomes_ignore) {
+    includeConfig 'conf/igenomes.config'
+} else {
+    params.genomes = [:]
+}
+
+
+// Export these variables to prevent local Python/R libraries from conflicting with those in the container
+// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container.
+// See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable.
+
+env {
+    PYTHONNOUSERSITE = 1
+    R_PROFILE_USER   = "/.Rprofile"
+    R_ENVIRON_USER   = "/.Renviron"
+    JULIA_DEPOT_PATH = "/usr/local/share/julia"
+}
+
+// Capture exit codes from upstream processes when piping
+process.shell = ['/bin/bash', '-euo', 'pipefail']
+
+def trace_timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss')
+timeline {
+    enabled = true
+    file    = "${params.tracedir}/execution_timeline_${trace_timestamp}.html"
+}
+report {
+    enabled = true
+    file    = "${params.tracedir}/execution_report_${trace_timestamp}.html"
+}
+trace {
+    enabled = true
+    file    = "${params.tracedir}/execution_trace_${trace_timestamp}.txt"
+}
+dag {
+    enabled = true
+    file    = "${params.tracedir}/pipeline_dag_${trace_timestamp}.html"
+}
+
+manifest {
+    name            = 'nf-core/rnadeseq'
+    author          = """"""
+    homePage        = 'https://github.com/nf-core/rnadeseq'
+    description     = """"""
+    mainScript      = 'main.nf'
+    nextflowVersion = '!>=22.10.1'
+    version         = '2.1dev'
+    doi             = ''
+}
+
+// Load modules.config for DSL2 module specific options
+includeConfig 'conf/modules.config'
+
+// Function to ensure that resource requirements don't go beyond
+// a maximum limit
+def check_max(obj, type) {
+    if (type == 'memory') {
+        try {
+            if (obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1)
+                return params.max_memory as nextflow.util.MemoryUnit
+            else
+                return obj
+        } catch (all) {
+            println "   ### ERROR ###   Max memory '${params.max_memory}' is not valid! Using default value: $obj"
+            return obj
+        }
+    } else if (type == 'time') {
+        try {
+            if (obj.compareTo(params.max_time as nextflow.util.Duration) == 1)
+                return params.max_time as nextflow.util.Duration
+            else
+                return obj
+        } catch (all) {
+            println "   ### ERROR ###   Max time '${params.max_time}' is not valid! Using default value: $obj"
+            return obj
+        }
+    } else if (type == 'cpus') {
+        try {
+            return Math.min( obj, params.max_cpus as int )
+        } catch (all) {
+            println "   ### ERROR ###   Max cpus '${params.max_cpus}' is not valid! Using default value: $obj"
+            return obj
+        }
+    }
+}
diff --git a/nextflow_schema.json b/nextflow_schema.json
new file mode 100644
index 00000000..8b0e0d4e
--- /dev/null
+++ b/nextflow_schema.json
@@ -0,0 +1,287 @@
+{
+    "$schema": "http://json-schema.org/draft-07/schema",
+    "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/nextflow_schema.json",
+    "title": "nf-core/rnadeseq pipeline parameters",
+    "description": "",
+    "type": "object",
+    "definitions": {
+        "input_output_options": {
+            "title": "Input/output options",
+            "type": "object",
+            "fa_icon": "fas fa-terminal",
+            "description": "Define where the pipeline should find input data and save output data.",
+            "required": ["input", "outdir"],
+            "properties": {
+                "input": {
+                    "type": "string",
+                    "format": "file-path",
+                    "mimetype": "text/csv",
+                    "pattern": "^\\S+\\.csv$",
+                    "schema": "assets/schema_input.json",
+                    "description": "Path to comma-separated file containing information about the samples in the experiment.",
+                    "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row. See [usage docs](https://nf-co.re/rnadeseq/usage#samplesheet-input).",
+                    "fa_icon": "fas fa-file-csv"
+                },
+                "outdir": {
+                    "type": "string",
+                    "format": "directory-path",
+                    "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.",
+                    "fa_icon": "fas fa-folder-open"
+                },
+                "email": {
+                    "type": "string",
+                    "description": "Email address for completion summary.",
+                    "fa_icon": "fas fa-envelope",
+                    "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.",
+                    "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$"
+                },
+                "multiqc_title": {
+                    "type": "string",
+                    "description": "MultiQC report title. Printed as page header, used for filename if not otherwise specified.",
+                    "fa_icon": "fas fa-file-signature"
+                }
+            }
+        },
+        "reference_genome_options": {
+            "title": "Reference genome options",
+            "type": "object",
+            "fa_icon": "fas fa-dna",
+            "description": "Reference genome related files and options required for the workflow.",
+            "properties": {
+                "genome": {
+                    "type": "string",
+                    "description": "Name of iGenomes reference.",
+                    "fa_icon": "fas fa-book",
+                    "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files e.g. `--genome GRCh38`. \n\nSee the [nf-core website docs](https://nf-co.re/usage/reference_genomes) for more details."
+                },
+                "fasta": {
+                    "type": "string",
+                    "format": "file-path",
+                    "mimetype": "text/plain",
+                    "pattern": "^\\S+\\.fn?a(sta)?(\\.gz)?$",
+                    "description": "Path to FASTA genome file.",
+                    "help_text": "This parameter is *mandatory* if `--genome` is not specified. If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs.",
+                    "fa_icon": "far fa-file-code"
+                },
+                "igenomes_base": {
+                    "type": "string",
+                    "format": "directory-path",
+                    "description": "Directory / URL base for iGenomes references.",
+                    "default": "s3://ngi-igenomes/igenomes",
+                    "fa_icon": "fas fa-cloud-download-alt",
+                    "hidden": true
+                },
+                "igenomes_ignore": {
+                    "type": "boolean",
+                    "description": "Do not load the iGenomes reference config.",
+                    "fa_icon": "fas fa-ban",
+                    "hidden": true,
+                    "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`."
+                }
+            }
+        },
+        "institutional_config_options": {
+            "title": "Institutional config options",
+            "type": "object",
+            "fa_icon": "fas fa-university",
+            "description": "Parameters used to describe centralised config profiles. These should not be edited.",
+            "help_text": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.",
+            "properties": {
+                "custom_config_version": {
+                    "type": "string",
+                    "description": "Git commit id for Institutional configs.",
+                    "default": "master",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "custom_config_base": {
+                    "type": "string",
+                    "description": "Base directory for Institutional configs.",
+                    "default": "https://raw.githubusercontent.com/nf-core/configs/master",
+                    "hidden": true,
+                    "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.",
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_name": {
+                    "type": "string",
+                    "description": "Institutional config name.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_description": {
+                    "type": "string",
+                    "description": "Institutional config description.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_contact": {
+                    "type": "string",
+                    "description": "Institutional config contact information.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_url": {
+                    "type": "string",
+                    "description": "Institutional config URL link.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                }
+            }
+        },
+        "max_job_request_options": {
+            "title": "Max job request options",
+            "type": "object",
+            "fa_icon": "fab fa-acquisitions-incorporated",
+            "description": "Set the top limit for requested resources for any single job.",
+            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
+            "properties": {
+                "max_cpus": {
+                    "type": "integer",
+                    "description": "Maximum number of CPUs that can be requested for any single job.",
+                    "default": 16,
+                    "fa_icon": "fas fa-microchip",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`"
+                },
+                "max_memory": {
+                    "type": "string",
+                    "description": "Maximum amount of memory that can be requested for any single job.",
+                    "default": "128.GB",
+                    "fa_icon": "fas fa-memory",
+                    "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`"
+                },
+                "max_time": {
+                    "type": "string",
+                    "description": "Maximum amount of time that can be requested for any single job.",
+                    "default": "240.h",
+                    "fa_icon": "far fa-clock",
+                    "pattern": "^(\\d+\\.?\\s*(s|m|h|day)\\s*)+$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`"
+                }
+            }
+        },
+        "generic_options": {
+            "title": "Generic options",
+            "type": "object",
+            "fa_icon": "fas fa-file-import",
+            "description": "Less common options for the pipeline, typically set in a config file.",
+            "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.",
+            "properties": {
+                "help": {
+                    "type": "boolean",
+                    "description": "Display help text.",
+                    "fa_icon": "fas fa-question-circle",
+                    "hidden": true
+                },
+                "version": {
+                    "type": "boolean",
+                    "description": "Display version and exit.",
+                    "fa_icon": "fas fa-question-circle",
+                    "hidden": true
+                },
+                "publish_dir_mode": {
+                    "type": "string",
+                    "default": "copy",
+                    "description": "Method used to save pipeline results to output directory.",
+                    "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.",
+                    "fa_icon": "fas fa-copy",
+                    "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"],
+                    "hidden": true
+                },
+                "email_on_fail": {
+                    "type": "string",
+                    "description": "Email address for completion summary, only when pipeline fails.",
+                    "fa_icon": "fas fa-exclamation-triangle",
+                    "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$",
+                    "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.",
+                    "hidden": true
+                },
+                "plaintext_email": {
+                    "type": "boolean",
+                    "description": "Send plain-text email instead of HTML.",
+                    "fa_icon": "fas fa-remove-format",
+                    "hidden": true
+                },
+                "max_multiqc_email_size": {
+                    "type": "string",
+                    "description": "File size limit when attaching MultiQC reports to summary emails.",
+                    "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$",
+                    "default": "25.MB",
+                    "fa_icon": "fas fa-file-upload",
+                    "hidden": true
+                },
+                "monochrome_logs": {
+                    "type": "boolean",
+                    "description": "Do not use coloured log outputs.",
+                    "fa_icon": "fas fa-palette",
+                    "hidden": true
+                },
+                "hook_url": {
+                    "type": "string",
+                    "description": "Incoming hook URL for messaging service",
+                    "fa_icon": "fas fa-people-group",
+                    "help_text": "Incoming hook URL for messaging service. Currently, MS Teams and Slack are supported.",
+                    "hidden": true
+                },
+                "multiqc_config": {
+                    "type": "string",
+                    "description": "Custom config file to supply to MultiQC.",
+                    "fa_icon": "fas fa-cog",
+                    "hidden": true
+                },
+                "multiqc_logo": {
+                    "type": "string",
+                    "description": "Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file",
+                    "fa_icon": "fas fa-image",
+                    "hidden": true
+                },
+                "multiqc_methods_description": {
+                    "type": "string",
+                    "description": "Custom MultiQC yaml file containing HTML including a methods description.",
+                    "fa_icon": "fas fa-cog"
+                },
+                "tracedir": {
+                    "type": "string",
+                    "description": "Directory to keep pipeline Nextflow logs and reports.",
+                    "default": "${params.outdir}/pipeline_info",
+                    "fa_icon": "fas fa-cogs",
+                    "hidden": true
+                },
+                "validate_params": {
+                    "type": "boolean",
+                    "description": "Boolean whether to validate parameters against the schema at runtime",
+                    "default": true,
+                    "fa_icon": "fas fa-check-square",
+                    "hidden": true
+                },
+                "show_hidden_params": {
+                    "type": "boolean",
+                    "fa_icon": "far fa-eye-slash",
+                    "description": "Show all params when using `--help`",
+                    "hidden": true,
+                    "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters."
+                }
+            }
+        }
+    },
+    "allOf": [
+        {
+            "$ref": "#/definitions/input_output_options"
+        },
+        {
+            "$ref": "#/definitions/reference_genome_options"
+        },
+        {
+            "$ref": "#/definitions/institutional_config_options"
+        },
+        {
+            "$ref": "#/definitions/max_job_request_options"
+        },
+        {
+            "$ref": "#/definitions/generic_options"
+        }
+    ]
+}
diff --git a/pyproject.toml b/pyproject.toml
new file mode 100644
index 00000000..0d62beb6
--- /dev/null
+++ b/pyproject.toml
@@ -0,0 +1,10 @@
+# Config file for Python. Mostly used to configure linting of bin/check_samplesheet.py with Black.
+# Should be kept the same as nf-core/tools to avoid fighting with template synchronisation.
+[tool.black]
+line-length = 120
+target_version = ["py37", "py38", "py39", "py310"]
+
+[tool.isort]
+profile = "black"
+known_first_party = ["nf_core"]
+multi_line_output = 3
diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf
new file mode 100644
index 00000000..0aecf87f
--- /dev/null
+++ b/subworkflows/local/input_check.nf
@@ -0,0 +1,44 @@
+//
+// Check input samplesheet and get read channels
+//
+
+include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check'
+
+workflow INPUT_CHECK {
+    take:
+    samplesheet // file: /path/to/samplesheet.csv
+
+    main:
+    SAMPLESHEET_CHECK ( samplesheet )
+        .csv
+        .splitCsv ( header:true, sep:',' )
+        .map { create_fastq_channel(it) }
+        .set { reads }
+
+    emit:
+    reads                                     // channel: [ val(meta), [ reads ] ]
+    versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ]
+}
+
+// Function to get list of [ meta, [ fastq_1, fastq_2 ] ]
+def create_fastq_channel(LinkedHashMap row) {
+    // create meta map
+    def meta = [:]
+    meta.id         = row.sample
+    meta.single_end = row.single_end.toBoolean()
+
+    // add path(s) of the fastq file(s) to the meta map
+    def fastq_meta = []
+    if (!file(row.fastq_1).exists()) {
+        exit 1, "ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}"
+    }
+    if (meta.single_end) {
+        fastq_meta = [ meta, [ file(row.fastq_1) ] ]
+    } else {
+        if (!file(row.fastq_2).exists()) {
+            exit 1, "ERROR: Please check input samplesheet -> Read 2 FastQ file does not exist!\n${row.fastq_2}"
+        }
+        fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ]
+    }
+    return fastq_meta
+}
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
new file mode 100644
index 00000000..6cdfcc1b
--- /dev/null
+++ b/workflows/rnadeseq.nf
@@ -0,0 +1,132 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    VALIDATE INPUTS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+def summary_params = NfcoreSchema.paramsSummaryMap(workflow, params)
+
+// Validate input parameters
+WorkflowRnadeseq.initialise(params, log)
+
+// TODO nf-core: Add all file path parameters for the pipeline to the list below
+// Check input path parameters to see if they exist
+def checkPathParamList = [ params.input, params.multiqc_config, params.fasta ]
+for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } }
+
+// Check mandatory parameters
+if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input samplesheet not specified!' }
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    CONFIG FILES
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+ch_multiqc_config          = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true)
+ch_multiqc_custom_config   = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty()
+ch_multiqc_logo            = params.multiqc_logo   ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty()
+ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true)
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    IMPORT LOCAL MODULES/SUBWORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// SUBWORKFLOW: Consisting of a mix of local and nf-core/modules
+//
+include { INPUT_CHECK } from '../subworkflows/local/input_check'
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    IMPORT NF-CORE MODULES/SUBWORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// MODULE: Installed directly from nf-core/modules
+//
+include { FASTQC                      } from '../modules/nf-core/fastqc/main'
+include { MULTIQC                     } from '../modules/nf-core/multiqc/main'
+include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main'
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    RUN MAIN WORKFLOW
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+// Info required for completion email and summary
+def multiqc_report = []
+
+workflow RNADESEQ {
+
+    ch_versions = Channel.empty()
+
+    //
+    // SUBWORKFLOW: Read in samplesheet, validate and stage input files
+    //
+    INPUT_CHECK (
+        ch_input
+    )
+    ch_versions = ch_versions.mix(INPUT_CHECK.out.versions)
+
+    //
+    // MODULE: Run FastQC
+    //
+    FASTQC (
+        INPUT_CHECK.out.reads
+    )
+    ch_versions = ch_versions.mix(FASTQC.out.versions.first())
+
+    CUSTOM_DUMPSOFTWAREVERSIONS (
+        ch_versions.unique().collectFile(name: 'collated_versions.yml')
+    )
+
+    //
+    // MODULE: MultiQC
+    //
+    workflow_summary    = WorkflowRnadeseq.paramsSummaryMultiqc(workflow, summary_params)
+    ch_workflow_summary = Channel.value(workflow_summary)
+
+    methods_description    = WorkflowRnadeseq.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description)
+    ch_methods_description = Channel.value(methods_description)
+
+    ch_multiqc_files = Channel.empty()
+    ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml'))
+    ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml'))
+    ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect())
+    ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([]))
+
+    MULTIQC (
+        ch_multiqc_files.collect(),
+        ch_multiqc_config.toList(),
+        ch_multiqc_custom_config.toList(),
+        ch_multiqc_logo.toList()
+    )
+    multiqc_report = MULTIQC.out.report.toList()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    COMPLETION EMAIL AND SUMMARY
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+workflow.onComplete {
+    if (params.email || params.email_on_fail) {
+        NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report)
+    }
+    NfcoreTemplate.summary(workflow, params, log)
+    if (params.hook_url) {
+        NfcoreTemplate.IM_notification(workflow, params, summary_params, projectDir, log)
+    }
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    THE END
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/

From ef41c6e58a4dedb1a57c15b06c98c57c7740a542 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 11:36:18 +0200
Subject: [PATCH 040/124] updated md5sums

---
 modules.json               | 10 ++++++++++
 tests/test.yml             | 10 +++++-----
 tests/test_full.yml        | 10 +++++-----
 tests/test_star_salmon.yml |  2 +-
 4 files changed, 21 insertions(+), 11 deletions(-)

diff --git a/modules.json b/modules.json
index dcb02c6f..1c3230cf 100644
--- a/modules.json
+++ b/modules.json
@@ -9,6 +9,16 @@
                         "branch": "master",
                         "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
                         "installed_by": ["modules"]
+                    },
+                    "fastqc": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
+                    },
+                    "multiqc": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
                     }
                 }
             }
diff --git a/tests/test.yml b/tests/test.yml
index 7ed85404..4b9b6105 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -66,11 +66,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: 237e9630d2998badcc69b1e193d5e1ab
+      md5sum: be0f292a205f2ecd453ee27ecdc62615
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 7ee7cd938140217a863774c3689ff1af
+      md5sum: f0e6d8117e3ff05ddabc3abb98f514b7
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 18873d39d839477337312cce0ec63398
+      md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf
@@ -82,9 +82,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 9318d57b86ba653f6183e61840a7943b
+      md5sum: 0bd6dfd32bad6f5292f6cde91f019844
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: d5632f9240116f42de59f05b7e822a29
+      md5sum: 452ca5dd8d5f190648988502f85d3ef1
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg
diff --git a/tests/test_full.yml b/tests/test_full.yml
index d739d687..77abaeee 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -63,11 +63,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv
-      md5sum: 237e9630d2998badcc69b1e193d5e1ab
+      md5sum: be0f292a205f2ecd453ee27ecdc62615
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv
-      md5sum: 7ee7cd938140217a863774c3689ff1af
+      md5sum: f0e6d8117e3ff05ddabc3abb98f514b7
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv
-      md5sum: 18873d39d839477337312cce0ec63398
+      md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf
@@ -76,9 +76,9 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png
     - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv
-      md5sum: 9318d57b86ba653f6183e61840a7943b
+      md5sum: 0bd6dfd32bad6f5292f6cde91f019844
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv
-      md5sum: d5632f9240116f42de59f05b7e822a29
+      md5sum: 452ca5dd8d5f190648988502f85d3ef1
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index a53cd1fd..bc2db1bb 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -70,7 +70,7 @@
     - path: results_test/differential_gene_expression/plots/PCA_plot.png
     - path: results_test/differential_gene_expression/plots/PCA_plot.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 710bb4810bcf109e03b559cae54d5031
+      md5sum: cd967cc355045d9dc8c0bb6ffcf40b61
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
       md5sum: 68b329da9893e34099c7d8ad5cb9c940
     - path: results_test/RNAseq_report.html

From 63525edfda80945dcc597b55a35919e923363777 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 11:59:15 +0200
Subject: [PATCH 041/124] updated rsem 5sums

---
 tests/test_star_rsem.yml | 12 +++++-------
 1 file changed, 5 insertions(+), 7 deletions(-)

diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index 377b99a8..e55c497e 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -88,11 +88,11 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 7f71760d73cab3c0d76a59c5ecdd78c0
+      md5sum: b1c0a1374c963fff71083a9cdefe8e9b
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 2b2ae0c1d4b4de442ced8d37193f61d4
+      md5sum: 631e89b4de9042936fa2652bde036e7d
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: a5422d271a102dff6c14ecfa46abdeab
+      md5sum: 80123c4ba2c97eaa0217816f41122855
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf
@@ -106,9 +106,7 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv
-      md5sum: 210529cfe4281b056e5bd59550757295
-    - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv
-      md5sum: 2b2ae0c1d4b4de442ced8d37193f61d4
+      md5sum: 276e871c81e85217bbd7eef951676084
     - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv
-      md5sum: dd87833db2f333dd2f9193c84edf23e5
+      md5sum: 9e6847fe5d631f09305dfa00dd3472d1
     - path: results_test/RNAseq_report.html

From e255f96465e9cee48acc1616bb24061f786f4609 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 12:00:41 +0200
Subject: [PATCH 042/124] Linting, prettier

---
 .nf-core.yml              | 2 +-
 lib/NfcoreSchema.groovy   | 4 +++-
 lib/NfcoreTemplate.groovy | 5 -----
 3 files changed, 4 insertions(+), 7 deletions(-)

diff --git a/.nf-core.yml b/.nf-core.yml
index 82415de5..a797f271 100644
--- a/.nf-core.yml
+++ b/.nf-core.yml
@@ -36,4 +36,4 @@ lint:
     - show_hidden_params
 
   multiqc_config:
-    - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'.                                                                           │
\ No newline at end of file
+    - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'.                                                                           │
diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy
index 33cd4f6e..9b34804d 100755
--- a/lib/NfcoreSchema.groovy
+++ b/lib/NfcoreSchema.groovy
@@ -2,6 +2,7 @@
 // This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template.
 //
 
+import nextflow.Nextflow
 import org.everit.json.schema.Schema
 import org.everit.json.schema.loader.SchemaLoader
 import org.everit.json.schema.ValidationException
@@ -83,6 +84,7 @@ class NfcoreSchema {
             'stub-run',
             'test',
             'w',
+            'with-apptainer',
             'with-charliecloud',
             'with-conda',
             'with-dag',
@@ -177,7 +179,7 @@ class NfcoreSchema {
         }
 
         if (has_error) {
-            System.exit(1)
+            Nextflow.error('Exiting!')
         }
     }
 
diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
index 25a0a74a..2023f451 100755
--- a/lib/NfcoreTemplate.groovy
+++ b/lib/NfcoreTemplate.groovy
@@ -323,11 +323,6 @@ class NfcoreTemplate {
         String.format(
             """\n
             ${dashedLine(monochrome_logs)}
-                                                    ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset}
-            ${colors.blue}        ___     __   __   __   ___     ${colors.green}/,-._.--~\'${colors.reset}
-            ${colors.blue}  |\\ | |__  __ /  ` /  \\ |__) |__         ${colors.yellow}}  {${colors.reset}
-            ${colors.blue}  | \\| |       \\__, \\__/ |  \\ |___     ${colors.green}\\`-._,-`-,${colors.reset}
-                                                    ${colors.green}`._,._,\'${colors.reset}
             ${colors.purple}  ${workflow.manifest.name} ${workflow_version}${colors.reset}
             ${dashedLine(monochrome_logs)}
             """.stripIndent()

From 578c8203b1facf2f342a6523bfee3ed1178d4b72 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 12:25:44 +0200
Subject: [PATCH 043/124] Updated nfcoreschema

---
 lib/NfcoreSchema.groovy | 4 +---
 1 file changed, 1 insertion(+), 3 deletions(-)

diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy
index 9b34804d..33cd4f6e 100755
--- a/lib/NfcoreSchema.groovy
+++ b/lib/NfcoreSchema.groovy
@@ -2,7 +2,6 @@
 // This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template.
 //
 
-import nextflow.Nextflow
 import org.everit.json.schema.Schema
 import org.everit.json.schema.loader.SchemaLoader
 import org.everit.json.schema.ValidationException
@@ -84,7 +83,6 @@ class NfcoreSchema {
             'stub-run',
             'test',
             'w',
-            'with-apptainer',
             'with-charliecloud',
             'with-conda',
             'with-dag',
@@ -179,7 +177,7 @@ class NfcoreSchema {
         }
 
         if (has_error) {
-            Nextflow.error('Exiting!')
+            System.exit(1)
         }
     }
 

From 1b69cb341b71854bde4be293f8a3968a7717634d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 13:12:34 +0200
Subject: [PATCH 044/124] removed redundant modules, corrected pipeline version

---
 CHANGELOG.md                     |  2 +-
 assets/samplesheet.csv           |  3 --
 modules.json                     | 10 ------
 modules/nf-core/fastqc/main.nf   | 51 -----------------------------
 modules/nf-core/fastqc/meta.yml  | 52 ------------------------------
 modules/nf-core/multiqc/main.nf  | 53 ------------------------------
 modules/nf-core/multiqc/meta.yml | 55 --------------------------------
 nextflow.config                  |  2 +-
 8 files changed, 2 insertions(+), 226 deletions(-)
 delete mode 100644 assets/samplesheet.csv
 delete mode 100644 modules/nf-core/fastqc/main.nf
 delete mode 100644 modules/nf-core/fastqc/meta.yml
 delete mode 100644 modules/nf-core/multiqc/main.nf
 delete mode 100644 modules/nf-core/multiqc/meta.yml

diff --git a/CHANGELOG.md b/CHANGELOG.md
index c455095d..48990c0c 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -3,7 +3,7 @@
 The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
 and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
 
-## 2.1dev
+## 2.2dev
 
 ### Added
 
diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv
deleted file mode 100644
index 5f653ab7..00000000
--- a/assets/samplesheet.csv
+++ /dev/null
@@ -1,3 +0,0 @@
-sample,fastq_1,fastq_2
-SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz
-SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz,
diff --git a/modules.json b/modules.json
index 1c3230cf..dcb02c6f 100644
--- a/modules.json
+++ b/modules.json
@@ -9,16 +9,6 @@
                         "branch": "master",
                         "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
                         "installed_by": ["modules"]
-                    },
-                    "fastqc": {
-                        "branch": "master",
-                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
-                        "installed_by": ["modules"]
-                    },
-                    "multiqc": {
-                        "branch": "master",
-                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
-                        "installed_by": ["modules"]
                     }
                 }
             }
diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf
deleted file mode 100644
index 9ae58381..00000000
--- a/modules/nf-core/fastqc/main.nf
+++ /dev/null
@@ -1,51 +0,0 @@
-process FASTQC {
-    tag "$meta.id"
-    label 'process_medium'
-
-    conda "bioconda::fastqc=0.11.9"
-    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
-        'https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0' :
-        'quay.io/biocontainers/fastqc:0.11.9--0' }"
-
-    input:
-    tuple val(meta), path(reads)
-
-    output:
-    tuple val(meta), path("*.html"), emit: html
-    tuple val(meta), path("*.zip") , emit: zip
-    path  "versions.yml"           , emit: versions
-
-    when:
-    task.ext.when == null || task.ext.when
-
-    script:
-    def args = task.ext.args ?: ''
-    def prefix = task.ext.prefix ?: "${meta.id}"
-    // Make list of old name and new name pairs to use for renaming in the bash while loop
-    def old_new_pairs = reads instanceof Path || reads.size() == 1 ? [[ reads, "${prefix}.${reads.extension}" ]] : reads.withIndex().collect { entry, index -> [ entry, "${prefix}_${index + 1}.${entry.extension}" ] }
-    def rename_to = old_new_pairs*.join(' ').join(' ')
-    def renamed_files = old_new_pairs.collect{ old_name, new_name -> new_name }.join(' ')
-    """
-    printf "%s %s\\n" $rename_to | while read old_name new_name; do
-        [ -f "\${new_name}" ] || ln -s \$old_name \$new_name
-    done
-    fastqc $args --threads $task.cpus $renamed_files
-
-    cat <<-END_VERSIONS > versions.yml
-    "${task.process}":
-        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
-    END_VERSIONS
-    """
-
-    stub:
-    def prefix = task.ext.prefix ?: "${meta.id}"
-    """
-    touch ${prefix}.html
-    touch ${prefix}.zip
-
-    cat <<-END_VERSIONS > versions.yml
-    "${task.process}":
-        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
-    END_VERSIONS
-    """
-}
diff --git a/modules/nf-core/fastqc/meta.yml b/modules/nf-core/fastqc/meta.yml
deleted file mode 100644
index 4da5bb5a..00000000
--- a/modules/nf-core/fastqc/meta.yml
+++ /dev/null
@@ -1,52 +0,0 @@
-name: fastqc
-description: Run FastQC on sequenced reads
-keywords:
-  - quality control
-  - qc
-  - adapters
-  - fastq
-tools:
-  - fastqc:
-      description: |
-        FastQC gives general quality metrics about your reads.
-        It provides information about the quality score distribution
-        across your reads, the per base sequence content (%A/C/G/T).
-        You get information about adapter contamination and other
-        overrepresented sequences.
-      homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/
-      documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/
-      licence: ["GPL-2.0-only"]
-input:
-  - meta:
-      type: map
-      description: |
-        Groovy Map containing sample information
-        e.g. [ id:'test', single_end:false ]
-  - reads:
-      type: file
-      description: |
-        List of input FastQ files of size 1 and 2 for single-end and paired-end data,
-        respectively.
-output:
-  - meta:
-      type: map
-      description: |
-        Groovy Map containing sample information
-        e.g. [ id:'test', single_end:false ]
-  - html:
-      type: file
-      description: FastQC report
-      pattern: "*_{fastqc.html}"
-  - zip:
-      type: file
-      description: FastQC report archive
-      pattern: "*_{fastqc.zip}"
-  - versions:
-      type: file
-      description: File containing software versions
-      pattern: "versions.yml"
-authors:
-  - "@drpatelh"
-  - "@grst"
-  - "@ewels"
-  - "@FelixKrueger"
diff --git a/modules/nf-core/multiqc/main.nf b/modules/nf-core/multiqc/main.nf
deleted file mode 100644
index 68f66bea..00000000
--- a/modules/nf-core/multiqc/main.nf
+++ /dev/null
@@ -1,53 +0,0 @@
-process MULTIQC {
-    label 'process_single'
-
-    conda "bioconda::multiqc=1.13"
-    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
-        'https://depot.galaxyproject.org/singularity/multiqc:1.13--pyhdfd78af_0' :
-        'quay.io/biocontainers/multiqc:1.13--pyhdfd78af_0' }"
-
-    input:
-    path  multiqc_files, stageAs: "?/*"
-    path(multiqc_config)
-    path(extra_multiqc_config)
-    path(multiqc_logo)
-
-    output:
-    path "*multiqc_report.html", emit: report
-    path "*_data"              , emit: data
-    path "*_plots"             , optional:true, emit: plots
-    path "versions.yml"        , emit: versions
-
-    when:
-    task.ext.when == null || task.ext.when
-
-    script:
-    def args = task.ext.args ?: ''
-    def config = multiqc_config ? "--config $multiqc_config" : ''
-    def extra_config = extra_multiqc_config ? "--config $extra_multiqc_config" : ''
-    """
-    multiqc \\
-        --force \\
-        $args \\
-        $config \\
-        $extra_config \\
-        .
-
-    cat <<-END_VERSIONS > versions.yml
-    "${task.process}":
-        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
-    END_VERSIONS
-    """
-
-    stub:
-    """
-    touch multiqc_data
-    touch multiqc_plots
-    touch multiqc_report.html
-
-    cat <<-END_VERSIONS > versions.yml
-    "${task.process}":
-        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
-    END_VERSIONS
-    """
-}
diff --git a/modules/nf-core/multiqc/meta.yml b/modules/nf-core/multiqc/meta.yml
deleted file mode 100644
index ebc29b27..00000000
--- a/modules/nf-core/multiqc/meta.yml
+++ /dev/null
@@ -1,55 +0,0 @@
-name: MultiQC
-description: Aggregate results from bioinformatics analyses across many samples into a single report
-keywords:
-  - QC
-  - bioinformatics tools
-  - Beautiful stand-alone HTML report
-tools:
-  - multiqc:
-      description: |
-        MultiQC searches a given directory for analysis logs and compiles a HTML report.
-        It's a general use tool, perfect for summarising the output from numerous bioinformatics tools.
-      homepage: https://multiqc.info/
-      documentation: https://multiqc.info/docs/
-      licence: ["GPL-3.0-or-later"]
-
-input:
-  - multiqc_files:
-      type: file
-      description: |
-        List of reports / files recognised by MultiQC, for example the html and zip output of FastQC
-  - multiqc_config:
-      type: file
-      description: Optional config yml for MultiQC
-      pattern: "*.{yml,yaml}"
-  - extra_multiqc_config:
-      type: file
-      description: Second optional config yml for MultiQC. Will override common sections in multiqc_config.
-      pattern: "*.{yml,yaml}"
-  - multiqc_logo:
-      type: file
-      description: Optional logo file for MultiQC
-      pattern: "*.{png}"
-
-output:
-  - report:
-      type: file
-      description: MultiQC report file
-      pattern: "multiqc_report.html"
-  - data:
-      type: dir
-      description: MultiQC data dir
-      pattern: "multiqc_data"
-  - plots:
-      type: file
-      description: Plots created by MultiQC
-      pattern: "*_data"
-  - versions:
-      type: file
-      description: File containing software versions
-      pattern: "versions.yml"
-authors:
-  - "@abhi18av"
-  - "@bunop"
-  - "@drpatelh"
-  - "@jfy133"
diff --git a/nextflow.config b/nextflow.config
index efe3e3f5..a7cac8ee 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -216,7 +216,7 @@ manifest {
     description     = 'Differential gene expression analysis and pathway analysis of RNAseq data'
     mainScript      = 'main.nf'
     nextflowVersion = '!>=22.10.1'
-    version = '2.1dev'
+    version = '2.2dev'
 }
 
 // Load modules.config for DSL2 module specific options

From 8f0bbefc5b4fef39baeef71904238877550d9ebe Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 13:14:51 +0200
Subject: [PATCH 045/124] lint

---
 lib/NfcoreSchema.groovy | 4 +++-
 1 file changed, 3 insertions(+), 1 deletion(-)

diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy
index 33cd4f6e..9b34804d 100755
--- a/lib/NfcoreSchema.groovy
+++ b/lib/NfcoreSchema.groovy
@@ -2,6 +2,7 @@
 // This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template.
 //
 
+import nextflow.Nextflow
 import org.everit.json.schema.Schema
 import org.everit.json.schema.loader.SchemaLoader
 import org.everit.json.schema.ValidationException
@@ -83,6 +84,7 @@ class NfcoreSchema {
             'stub-run',
             'test',
             'w',
+            'with-apptainer',
             'with-charliecloud',
             'with-conda',
             'with-dag',
@@ -177,7 +179,7 @@ class NfcoreSchema {
         }
 
         if (has_error) {
-            System.exit(1)
+            Nextflow.error('Exiting!')
         }
     }
 

From 465618b2f89e5dc5030298cc5aa11fd9788a3e23 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 2 May 2023 13:56:37 +0200
Subject: [PATCH 046/124] changed python version in linting.yml

---
 .github/workflows/linting.yml | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml
index 858d622e..888cb4bc 100644
--- a/.github/workflows/linting.yml
+++ b/.github/workflows/linting.yml
@@ -78,7 +78,7 @@ jobs:
 
       - uses: actions/setup-python@v4
         with:
-          python-version: "3.7"
+          python-version: "3.8"
           architecture: "x64"
 
       - name: Install dependencies

From 7788bfa0c99ef0b66550ca7acf6db5f3818af325 Mon Sep 17 00:00:00 2001
From: Oskar Wacker <iivow01@rddsws.am10.uni-tuebingen.de>
Date: Wed, 14 Jun 2023 11:12:04 +0200
Subject: [PATCH 047/124] Added logic for transformation method

---
 assets/RNAseq_report.Rmd | 79 ++++++++++++++++++++++++----------------
 bin/Execute_report.R     |  5 ++-
 modules/local/report.nf  |  4 +-
 nextflow.config          |  2 +-
 nextflow_schema.json     |  9 +++--
 5 files changed, 59 insertions(+), 40 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 729a0af6..050a03e0 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -29,8 +29,8 @@ params:
     batch_effect: ''
     logFC_threshold: ''
     pval_threshold: ''
+    trafo: ''
     nsub_genes: ''
-    rlog: ''
     round_DE: ''
 
     #Pathway analysis args
@@ -89,7 +89,6 @@ label_size <- function(dimension_int) {
 ```
 
 ```{r param_validation, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
-
 #self-written isProvided function that tests for zero-length, emptystring and NULL (for some reason, some params cannot be tested with emptystring)
 isProvided <- function(value) {
     result <- TRUE
@@ -562,14 +561,42 @@ if (params$input_type == "rsem") {
 }
 
 ############################## TRANSFORMED AND NORMALIZED COUNTS ###################
-if (params$rlog){
+# Prepare variables and text snippets for report
+vst_cutoff <- 0.005
+var_sizeFactors <- var(sizeFactors(cds))
+run_rlog <- T
+trafo_long_text <- "regularized logarithm (rlog)"
+trafo_override_text <- ""
+nsub_genes_text <- ""
+if (params$trafo == "vst-force") {
+    run_rlog <- F
+    trafo_long_text <- "variant stabilizing transformation (vst)"
+    nsub_genes_text <- paste0("The number of subset genes for the vst transformation is ", as.character(params$nsub_genes), ".")
+    if (var_sizeFactors >= vst_cutoff) {
+        trafo_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
+        nsub_genes_text <- ""
+    }
+} else if (params$trafo == "vst") {
+    run_rlog <- F
+    trafo_long_text <- "variant stabilizing transformation (vst)"
+    nsub_genes_text <- paste0("The number of subset genes for the vst transformation is ", as.character(params$nsub_genes), ".")
+    if (var_sizeFactors >= vst_cutoff) {
+        run_rlog <- T
+        trafo_long_text <- "regularized logarithm (rlog)"
+        trafo_override_text <- paste0(" This was done as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
+        nsub_genes_text <- ""
+    }
+}
+trafo_text <- ifelse(run_rlog, "rlog", "vst")
+
+if (run_rlog){
 
     # rlog transformation
     rld <- rlog(cds, blind=FALSE)
     rld_names <- merge(x=gene_names, y=assay(rld), by.x = "Ensembl_ID", by.y="row.names")
     write.table(rld_names, "differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F,  qmethod = c("escape", "double"))
 
-    #save table to another variable if pathway analysis
+    # save table to another variable if pathway analysis
     if (params$pathway_analysis) {
         norm_counts <- rld_names
         rownames(norm_counts) <- norm_counts$Ensembl_ID
@@ -582,7 +609,7 @@ if (params$rlog){
     vsd_names <- merge(x=gene_names, y=assay(vsd), by.x = "Ensembl_ID", by.y="row.names")
     write.table(vsd_names, "differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double"))
 
-    #save table to another variable if pathway analysis
+    # save table to another variable if pathway analysis
     if (params$pathway_analysis) {
         norm_counts <- vsd_names
         rownames(norm_counts) <- norm_counts$Ensembl_ID
@@ -627,24 +654,21 @@ pdf("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_tra
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = "sd rlog transformed count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd ", if (params$rlog) "rld" else "vsd" ," transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
 dev.off()
 notAllZero <- (rowSums(counts(cds))>0)
 png("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png")
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = "sd rlog transformed count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd ", if (params$rlog) "rld" else "vsd" ," transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
 dev.off()
 notAllZero <- (rowSums(counts(cds))>0)
 svg("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg")
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = "sd rlog transformed count data")
-meanSdPlot(assay((if (params$rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd ", if (params$rlog) "rld" else "vsd" ," transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
 dev.off()
 
 # Further diagnostics plots
@@ -921,26 +945,19 @@ cat(paste0("***
 
 <!-- multiQC end -->
 
-```{r prepare_text}
-
-#Prepare text snippets
-rlog_text <- ifelse(params$rlog, "rlog", "vst")
-rlog_long_text <- ifelse(params$rlog, "regularized logarithm (rlog)", "variant stabilizing transformation (vst)")
-nsub_genes_text <- ifelse(params$rlog, "", paste("The number of subset genes for the 'vst' transformation is", as.character(params$nsub_genes), "."))
-```
-
 # Gene expression data
 
 ## Raw and normalized count tables
 
 The raw count table and normalized count tables are available [here](./differential_gene_expression/gene_counts_tables).
 The differential expression analysis is performed using the raw gene count table.
-For PCA analysis and heatmap plotting, the `r rlog_long_text` normalized gene counts were employed.
+For PCA analysis and heatmap plotting, the `r trafo_long_text` normalized gene counts were employed. 
+`r trafo_override_text`
 `r nsub_genes_text`
 
 ## Principal component analysis (PCA)
 
-A PCA plot of the `r rlog_text` normalized gene expression visualizes the clustering of samples according to their gene expression.
+A PCA plot of the `r trafo_text` normalized gene expression visualizes the clustering of samples according to their gene expression.
 In this way, the overall effect of experimental conditions and sample grouping, and any batch effects are visualized.
 The original plot can be downloaded [here](./differential_gene_expression/plots/PCA_plot.pdf).
 
@@ -948,7 +965,7 @@ The original plot can be downloaded [here](./differential_gene_expression/plots/
 
 ```{r PCA_ifNoBatch_plot, echo=F, message=F, warning=F, out.width="80%", dpi=1200, fig.align='center'}
 ############################ PCA PLOTS ########################
-pcaData <- plotPCA(if (params$rlog) rld else vsd,intgroup=c("combfactor"), ntop=dim(if (params$rlog) rld else vsd)[1], returnData=TRUE)
+pcaData <- plotPCA(if (run_rlog) rld else vsd,intgroup=c("combfactor"), ntop=dim(if (run_rlog) rld else vsd)[1], returnData=TRUE)
 percentVar <- round(100*attr(pcaData, "percentVar"))
 pca <- ggplot(pcaData, aes(PC1, PC2, color=combfactor, text=paste("Sample: ", name, "<br>", "PC1: ", PC1, "<br>", "PC2: ", PC2))) +
     geom_point(size=3) +
@@ -985,12 +1002,12 @@ A PCA of the batch-effect corrected data is shown below and can be found [here](
 ```
 
 ```{r PCA_ifBatch_plot, echo=F, message=F, warning=F, dpi=1200, fig.align='center', eval=params$batch_effect}
-if (params$rlog) {
+if (run_rlog) {
     assay(rld) <- limma::removeBatchEffect(assay(rld), rld$batch)
 } else {
     assay(vsd) <- limma::removeBatchEffect(assay(vsd), vsd$batch)
 }
-pcaData2 <- plotPCA(if (params$rlog) rld else vsd, intgroup=c("combfactor"), ntop = dim(if (params$rlog) rld else vsd)[1], returnData=TRUE)
+pcaData2 <- plotPCA(if (run_rlog) rld else vsd, intgroup=c("combfactor"), ntop = dim(if (run_rlog) rld else vsd)[1], returnData=TRUE)
 percentVar <- round(100*attr(pcaData, "percentVar"))
 pca2 <- ggplot(pcaData2, aes(PC1, PC2, color=combfactor, text=paste("Sample: ", name, "<br>", "PC1: ", PC1, "<br>", "PC2: ", PC2))) +
     geom_point(size=3)+
@@ -1031,7 +1048,7 @@ The original plot can be downloaded [here](./differential_gene_expression/plots/
 ```{r heatmap_calculate, include=FALSE}
 ##################  SAMPLE DISTANCES HEATMAP ##################
 # Sample distances, this chunk is hidden as otherwise, the heatmap is shown multiple times
-sampleDists <- dist(t(assay(if (params$rlog) rld else vsd)))
+sampleDists <- dist(t(assay(if (run_rlog) rld else vsd)))
 sampleDistMatrix <- as.matrix(sampleDists)
 colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255)
 par(oma=c(3,3,3,3))
@@ -1050,12 +1067,13 @@ dist_heatmap %>% layout(height=700, width=800) %>% config(modeBarButtonsToAdd =
 
 After unzipping the results ZIP file, the differential expression results can be found in the directory [differential_gene_expression/](./differential_gene_expression), containing the following subfolders:
 
-```{r aligner_strings, echo=FALSE, message=FALSE, warning=FALSE}
+```{r structure_strings, echo=FALSE, message=FALSE, warning=FALSE}
 
-# Change the different strings depending on which input_type was used
+# Change the different strings depending on which input_type and transformation was used
     featurecounts_string <- ifelse(params$input_type == "featurecounts", "
     - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline; used for the differential gene expression analysis.", "")
     rsem_salmon_string <- ifelse(params$input_type %in% c("rsem", "salmon"), paste0("- `star_", params$input_type, "_files.txt`: list of ", ifelse(params$input_type == "rsem", "RSEM counts files", "Salmon quantification files"), " that were provided as input; used for the differential gene expression analysis."), "")
+    genecounts_string <- ifelse(run_rlog, "- `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts.", "- `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation.")
 ```
 
 <br><details>
@@ -1068,14 +1086,13 @@ After unzipping the results ZIP file, the differential expression results can be
     `r rsem_salmon_string`
 - `gene_counts_tables/`
     `r featurecounts_string`
-    - `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts.
-    - `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation.
+    `r genecounts_string`
     - `sizeFactor_libraries.tsv`: size factors for each sample.
 - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast.
 - `final_gene_table/final_gene_list_DESeq2.tsv`: table containing a list of all genes considered in the analysis. Here a summary of the log2 Fold Change and p-adjusted values for all contrasts is displayed. Additionally, the column **filter** shows if this gene was differentially expressed (DE) in any of the contrasts, or not (not_DE). The column **contrast_vector** contains for each contrast considered in the analysis a 1 if the gene was differentially expressed for this contrast or a 0 if it was not.
 - `plots/`:
-    - `Heatmaps_of_distances.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the rlog normalized gene counts are considered.
-    - `PCA_plot.pdf`: PCA of the rlog normalized gene counts.
+    - `Heatmaps_of_distances.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r trafo_text` normalized gene counts are considered.
+    - `PCA_plot.pdf`: PCA of the `r trafo_text` normalized gene counts.
     - `boxplots_example_genes/`: boxplots of the normalized gene counts for each of the sample groups for example genes.
     - `boxplots_requested_genes/`: boxplots of the normalized gene counts for each of the sample groups for the list of requested genes.
     - `further_diagnostics_plots/`: plots for diagnostics of the differential gene analysis procedure.
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index c14dba59..a3573582 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -23,7 +23,8 @@ option_list = list(
     make_option("--pval_threshold", type="double", default=0.05, help="p value threshold to consider a gene DE."),
     make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
 
-    make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
+    #make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
+    make_option(c("-z", "--trafo"), type="character", default=NULL, help="Which transformation(s) to use."),
     make_option(c("-n", "--nsub_genes"), type="integer", default=NULL, help="Subset number of genes for vst."),
 
     make_option(c("-a", "--pathway_analysis"), action="store_true", default=FALSE, help="Whether to run pathway analysis."),
@@ -70,7 +71,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 batch_effect = opt$batch_effect,
                                 logFC_threshold = opt$logFC_threshold,
                                 pval_threshold = opt$pval_threshold,
-                                rlog = opt$rlog,
+                                trafo = opt$trafo,
                                 nsub_genes = opt$nsub_genes,
                                 round_DE = opt$round_DE,
 
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 1e6d8dc5..556ebad5 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -32,7 +32,7 @@ process REPORT {
     def genelist_opt = genelist.name != 'NO_FILE' ? "--genelist $genelist" : ''
     def relevel_opt = relevel.name != 'NO_FILE2' ? "--relevel $relevel" : ''
     def batch_effect_opt = params.batch_effect ? "--batch_effect TRUE" : ''
-    def rlog_opt = params.use_vst ? '--rlog FALSE' : ''
+//    def rlog_opt = params.use_vst ? '--rlog FALSE' : ''
     def round_DE_opt = params.round_DE ? "--round_DE $params.round_DE" : ''
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
@@ -67,7 +67,7 @@ process REPORT {
         $batch_effect_opt \
         --logFC_threshold $params.logFC_threshold \
         --pval_threshold $params.pval_threshold \
-        $rlog_opt \
+        --trafo $params.trafo \
         --nsub_genes $params.vst_genes_number \
         $round_DE_opt \
         $pathway_opt \
diff --git a/nextflow.config b/nextflow.config
index a7cac8ee..034f4d64 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -26,7 +26,7 @@ params {
     batch_effect = null
     logFC_threshold = 0
     pval_threshold = 0.05
-    use_vst = false
+    trafo = 'rlog'
     vst_genes_number = 1000
     round_DE = null
 
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 0f7a75cc..93d0fb07 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -80,10 +80,11 @@
                     "default": 0.05,
                     "description": "p value (float) to consider a gene as differentially expressed. The default value is 0.05."
                 },
-                "use_vst": {
-                    "type": "boolean",
-                    "default": false,
-                    "description": "Set this flag to true to use vst transformation (recommended when >50 samples; if false, will use rlog)"
+                "trafo": {
+                    "type": "string",
+                    "default": "rlog",
+                    "enum": ["rlog", "vst", "vst-force"],
+                    "description": "Which transformation to use during DE analysis; either rlog or vst or vst-force . If vst: the pipeline will override this and run rlog if it detects a strong variation in size factors in the input data (this will be stated in the report). To disable this behavior, set to vst-force."
                 },
                 "vst_genes_number": {
                     "type": "integer",

From 2cbb218dbf009992965b4f60342d729e640d2fc1 Mon Sep 17 00:00:00 2001
From: Oskar Wacker <iivow01@rddsws.am10.uni-tuebingen.de>
Date: Wed, 14 Jun 2023 15:01:44 +0200
Subject: [PATCH 048/124] updated changelog, changed text in report, updated
 docs

---
 CHANGELOG.md             | 1 +
 assets/RNAseq_report.Rmd | 4 ++--
 docs/usage.md            | 8 ++++----
 3 files changed, 7 insertions(+), 6 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 48990c0c..503fc46d 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst
 - [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
 - [#197](https://github.com/qbic-pipelines/rnadeseq/pull/197) Added gprofiler version string to report
 - [#196](https://github.com/qbic-pipelines/rnadeseq/pull/196) Added optional `--quote` parameter
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 050a03e0..321e6bfb 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -562,7 +562,7 @@ if (params$input_type == "rsem") {
 
 ############################## TRANSFORMED AND NORMALIZED COUNTS ###################
 # Prepare variables and text snippets for report
-vst_cutoff <- 0.005
+vst_cutoff <- 0.05
 var_sizeFactors <- var(sizeFactors(cds))
 run_rlog <- T
 trafo_long_text <- "regularized logarithm (rlog)"
@@ -1073,7 +1073,7 @@ After unzipping the results ZIP file, the differential expression results can be
     featurecounts_string <- ifelse(params$input_type == "featurecounts", "
     - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline; used for the differential gene expression analysis.", "")
     rsem_salmon_string <- ifelse(params$input_type %in% c("rsem", "salmon"), paste0("- `star_", params$input_type, "_files.txt`: list of ", ifelse(params$input_type == "rsem", "RSEM counts files", "Salmon quantification files"), " that were provided as input; used for the differential gene expression analysis."), "")
-    genecounts_string <- ifelse(run_rlog, "- `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts.", "- `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation.")
+    genecounts_string <- ifelse(run_rlog, '- `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts.', '- `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation.')
 ```
 
 <br><details>
diff --git a/docs/usage.md b/docs/usage.md
index 93002a3e..d7078cf1 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -30,7 +30,7 @@
   - [`--genelist`](#--genelist)
   - [`--batch_effect`](#--batch_effect)
   - [`--min_DEG_pathway`](#--min_deg_pathway)
-  - [`--use_vst`](#--use_vst)
+  - [`--trafo`](#--trafo)
   - [`--vst_genes_number`](#--vst_genes_number)
   - [`--round_DE`](#--round_DE)
   - [`--run_pathway_analysis`](#--run_pathway_analysis)
@@ -305,13 +305,13 @@ For more information, please check the [DESeq2 vignette](http://bioconductor.org
 
 Integer indicating how many genes in a pathway must be differentially expressed to be considered as enriched, and report these pathways in tables and the final report. The default value is 1.
 
-### `--use_vst`
+### `--trafo`
 
-Consider using this parameter when the number of input samples is greater than 50. With large input sample sizes the rlog transformation becomes very time consuming. Note: If this flag is used, the pathway analysis will make use of vst transformed counts instead of rlog transformed counts. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
+Set to `vst`, `vst-force` or `rlog` (default) to determine which transformation is applied to the data. If `vst`, the pipeline can still use rlog instead if the input data are found to have different size factor variances (>=0.05) which will be stated in the report. If you want to use the vst method regardless of data, use `vst-force`. Generally speaking, vst is faster but rlog is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
 
 ### `--vst_genes_number`
 
-This is ignored if `--use_vst` is set to false. If using the vst transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small rnaseq data. The default `vst` function for varianceStabilizingTransformation in DESeq2 is 1000, which triggers an error with small datasets. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
+This is ignored if `--trafo` is set to `rlog`. If using the vst transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small rnaseq data. The default `vst` function for varianceStabilizingTransformation in DESeq2 is 1000, which triggers an error with small datasets. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
 
 ### `--round_DE`
 

From 32ab7377289bbac5d9d2bd6b716a20be2d96a0e4 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 15 Jun 2023 14:16:44 +0200
Subject: [PATCH 049/124] checking if a fix is actually necessary

---
 assets/RNAseq_report.Rmd | 10 +++++-----
 1 file changed, 5 insertions(+), 5 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 779af63c..a0081176 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -140,15 +140,15 @@ isProvided <- function(value) {
 }
 
 #this function is used to produce a more informative error message when count table and metadata table disagree
-write_error_msg <- function(counts, meta) {
+write_error_msg <- function(counts, qbicCodes) {
     error_msg <- "Count table headers do not exactly match the metadata table sample names! Offending value(s) in count table:\n"
     for (c in names(counts)) {
-        if (!(c %in% row.names(meta))) {
+        if (!(c %in% qbicCodes)) {
             error_msg <- paste0(error_msg, c, "\n")
         }
     }
     error_msg <- paste0(error_msg, "\n", "Offending value(s) in metadata table:\n")
-    for (m in row.names(meta)) {
+    for (m in qbicCodes) {
         if (!(m %in% names(counts))) {
             error_msg <- paste0(error_msg, m, "\n")
         }
@@ -358,11 +358,11 @@ if (params$input_type == "featurecounts"){
 
     # Need to order columns in count.table
     count.table <- count.table[, order(names(count.table))]
-
     names(count.table) = metadata$sampleName
+
     if (!identical(names(count.table),row.names(metadata))) {
     print("After refactoring the metadata table, count table headers do not exactly match the metadata table sample names! Offending values in count table:")
-    error_out <- write_error_msg(count.table, metadata)
+    error_out <- write_error_msg(count.table, qbicCodes)
     stop(paste0(error_out, "\n"), call.=F)
     }
 

From 2c69782f8d64590bd39567c10590852a549adcbb Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 15 Jun 2023 15:46:36 +0200
Subject: [PATCH 050/124] cleaned up and rewrote error message

---
 assets/RNAseq_report.Rmd | 34 ++++++++++++++++++++++++++--------
 1 file changed, 26 insertions(+), 8 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index a0081176..2994f71c 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -141,19 +141,26 @@ isProvided <- function(value) {
 
 #this function is used to produce a more informative error message when count table and metadata table disagree
 write_error_msg <- function(counts, qbicCodes) {
-    error_msg <- "Count table headers do not exactly match the metadata table sample names! Offending value(s) in count table:\n"
+    error_msg <- "Count table headers do not exactly match the metadata table sample names!\n"
+    
+    counts_msg <- ""
+    counts_offending <- ""
     for (c in names(counts)) {
         if (!(c %in% qbicCodes)) {
-            error_msg <- paste0(error_msg, c, "\n")
+            counts_msg <- "Offending value(s) in count table:\n"
+            counts_offending <- paste0(counts_offending, c, "\n")
         }
     }
-    error_msg <- paste0(error_msg, "\n", "Offending value(s) in metadata table:\n")
+    
+    meta_msg <- ""
+    meta_offending <- ""
     for (m in qbicCodes) {
         if (!(m %in% names(counts))) {
-            error_msg <- paste0(error_msg, m, "\n")
+            meta_msg <- "Offending value(s) in metadata table:\n"
+            meta_offending <- paste0(meta_offending, m, "\n")
         }
     }
-    return(paste0(error_msg, "\n"))
+    return(paste0(error_msg, counts_msg, counts_offending, meta_msg, meta_offending))
 }
 
 #Check necessary params
@@ -358,13 +365,24 @@ if (params$input_type == "featurecounts"){
 
     # Need to order columns in count.table
     count.table <- count.table[, order(names(count.table))]
-    names(count.table) = metadata$sampleName
 
-    if (!identical(names(count.table),row.names(metadata))) {
-    print("After refactoring the metadata table, count table headers do not exactly match the metadata table sample names! Offending values in count table:")
+
+
+    
+    write(names(count.table), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/names_counttable")
+    write(metadata$sampleName, file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/metadata_sampleName")
+    write(row.names(metadata), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/rownames_metatable")
+    write(qbicCodes, file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/qbicCodes")
+    
+ 
+
+    if (!identical(names(count.table),qbicCodes)) {
     error_out <- write_error_msg(count.table, qbicCodes)
     stop(paste0(error_out, "\n"), call.=F)
     }
+   
+    names(count.table) = metadata$sampleName
+    write(names(count.table), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/names_counttable")
 
     # Write raw counts to file
     count_table_names <- merge(x=gene_names, y=count.table, by.x = "Ensembl_ID", by.y="row.names")

From 18560958c28e4038a506bdfc4c75e0077fccc53f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 15 Jun 2023 15:49:05 +0200
Subject: [PATCH 051/124] updated changelog

---
 CHANGELOG.md | 1 +
 1 file changed, 1 insertion(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 48990c0c..80b02e2b 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -21,6 +21,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Fixed
 
+- [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata
 - [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
 
 ## 2.1 - Wheat Fields

From cc63fbf6778c2176525c14a676e23b4a948ee342 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 15 Jun 2023 15:53:07 +0200
Subject: [PATCH 052/124] removed local write command

---
 assets/RNAseq_report.Rmd | 12 ------------
 1 file changed, 12 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 6563160f..e2758ca4 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -365,23 +365,11 @@ if (params$input_type == "featurecounts"){
     # Need to order columns in count.table
     count.table <- count.table[, order(names(count.table))]
 
-
-
-    
-    write(names(count.table), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/names_counttable")
-    write(metadata$sampleName, file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/metadata_sampleName")
-    write(row.names(metadata), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/rownames_metatable")
-    write(qbicCodes, file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/qbicCodes")
-    
- 
-
     if (!identical(names(count.table),qbicCodes)) {
     error_out <- write_error_msg(count.table, qbicCodes)
     stop(paste0(error_out, "\n"), call.=F)
     }
-   
     names(count.table) = metadata$sampleName
-    write(names(count.table), file="/sfs/9/ws/iivow01-test/rnadeseq/error_missing/names_counttable")
 
     # Write raw counts to file
     count_table_names <- merge(x=gene_names, y=count.table, by.x = "Ensembl_ID", by.y="row.names")

From d3ac03d6dd734f9356d1a10d940fcbf8e9729561 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 19 Jun 2023 10:08:11 +0200
Subject: [PATCH 053/124] Added separate sizefactor computation for tximport,
 changed < and > to <= and => for logFC and pval

---
 assets/RNAseq_report.Rmd | 59 +++++++++++++++++++++++-----------------
 conf/test_use_vst.config |  2 +-
 2 files changed, 35 insertions(+), 26 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 321e6bfb..b5bbd4db 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -55,8 +55,11 @@ params:
 # Author: Silvia Morini, Gisela Gabernet, Simon Heumos
 ---
 
-```{r seed_plot_peparations, include=F}
-#set a seed in case some background calculations are random-based
+```{r init, include=F}
+# The following envvar is necessary to enable <= and >= comparisons
+Sys.setenv("_R_USE_PIPEBIND_" = TRUE)
+
+# set a seed in case some background calculations are random-based
     set.seed(10)
 
 # svg path for the little svg format icon displayed in the interactive plots for svg export; based on https://www.svgviewer.dev/:
@@ -548,7 +551,13 @@ if (isProvided(params$path_relevel)) {
 }
 
 # SizeFactors(cds) as indicator of library sequencing depth
-write.table(sizeFactors(cds),paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T,  col.names = F, qmethod = c("escape", "double"))
+if (params$input_type == "featurecounts") {
+    sf <- sizeFactors(cds)
+} else {
+    nm <- assays(cds)[["avgTxLength"]]
+    sf <- estimateSizeFactorsForMatrix(counts(cds)/nm)
+}
+write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T,  col.names = F, qmethod = c("escape", "double"))
 
 # Write cds assay table to file
 write.table(counts(cds, normalized=T), paste("differential_gene_expression/gene_counts_tables/deseq2_table.tsv", sep=""), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
@@ -563,7 +572,7 @@ if (params$input_type == "rsem") {
 ############################## TRANSFORMED AND NORMALIZED COUNTS ###################
 # Prepare variables and text snippets for report
 vst_cutoff <- 0.05
-var_sizeFactors <- var(sizeFactors(cds))
+var_sizeFactors <- var(sf)
 run_rlog <- T
 trafo_long_text <- "regularized logarithm (rlog)"
 trafo_override_text <- ""
@@ -1151,7 +1160,7 @@ if (isProvided(params$path_contrast_matrix)){
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj < params$pval_threshold & abs(log2FoldChange) > params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1211,7 +1220,7 @@ if (isProvided(params$path_contrast_list)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj < params$pval_threshold & abs(log2FoldChange) > params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1253,7 +1262,7 @@ if (isProvided(params$path_contrast_pairs)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj < params$pval_threshold & abs(log2FoldChange) > params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
 
         # Round table before writing
@@ -1293,7 +1302,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj < params$pval_threshold & abs(log2FoldChange) > params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1326,11 +1335,11 @@ logFC = DE_genes_df[,logFC_cols,drop=F]
 padj = DE_genes_df[,padj_cols,drop=F]
 padj[is.na(padj)] <- 1
 
-# Convert to binary (1/0) matrix if padj < params$pval_threshold or not, respectively
-padj_bin = data.matrix(ifelse(padj < params$pval_threshold, 1, 0))
+# Convert to binary (1/0) matrix if padj <= params$pval_threshold or not, respectively
+padj_bin = data.matrix(ifelse(padj <= params$pval_threshold, 1, 0))
 
-# Convert to binary (1/0) matrix if logFC is bigger or smaller than threshold or not, respectively
-logFC_bin = data.matrix(ifelse(abs(logFC) > params$logFC_threshold, 1, 0))
+# Convert to binary (1/0) matrix if logFC is bigger/equal or smaller/equal than threshold or not, respectively
+logFC_bin = data.matrix(ifelse(abs(logFC) >= params$logFC_threshold, 1, 0))
 # Multiply the two bin matrices --> if padj matrix value or LogFC matrix value is 0, will be 0
 
 DE_bin = padj_bin * logFC_bin
@@ -1439,7 +1448,7 @@ log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10
 
 pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, "<br>", "Log2FC: ", formatC(log2FoldChange, digits=2)))) +
         #this goes after alpha=0.5:
-        geom_point(alpha=0.5, aes(color=ifelse(abs(DE_all$log2FoldChange) > params$logFC_threshold &  DE_all$logpval > -log10(params$pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
+        geom_point(alpha=0.5, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold &  DE_all$logpval >= -log10(params$pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
         geom_hline(yintercept = 16, linetype= "dashed", size = 0.2, color = "grey") +
         geom_hline(yintercept = -log10(params$pval_threshold), size = 0.2, color = "grey") +
         geom_vline(xintercept = -params$logFC_threshold, size = 0.2, color = "grey") +
@@ -2151,10 +2160,12 @@ as.character(params$revision),
 "` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `",
 quant_tool, "` were processed with the R package `DESeq2 v",
 packageVersion("DESeq2"),
-"` [@love2014differential]."
+"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", 
+pval_text, "for the p value and",
+logFC_text, "for the log2 Fold Change."
 ))
 
-```{r conditional_pathway, echo=FALSE, results='asis', eval=params$pathway_analysis&isProvided(params$path_software_versions)}
+```{r conditional_pathway, echo=FALSE, results='asis', eval=isProvided(params$pathway_analysis)&isProvided(params$path_software_versions)}
 version_gprofiler2 = packageVersion("gprofiler2")
 version_annotation = packageVersion("AnnotationDbi")
 name_species       = params$species_library
@@ -2175,16 +2186,14 @@ if (isProvided(params$custom_gmt)){
 }
 
 cat(paste0(
-"For pathway analysis, the R packages `gprofiler2 v",
-version_gprofiler2,
-" `, `AnnotationDbi v",
-version_annotation,
-"` and `",
-name_species,
-" v",
-version_annotation,
-"` were employed. ",
-database_string))
+"## Differential gene expression analysis
+
+For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2,
+" `, `AnnotationDbi v", version_annotation,
+"` and `", name_species, " v", version_annotation,
+"` were employed. ", database_string,
+"Pathways were classified as enriched for those genes with padj <= ", pval_text, "."
+))
 ```
 
 Graphs were produced in RStudio with ``r R.version.string`` mainly using the R package `ggplot2 v`r packageVersion("ggplot2")`` . Final reports were produced using the R package `rmarkdown v`r packageVersion("rmarkdown") ``, with the `knitr v`r packageVersion("knitr")`` and `DT v`r packageVersion("DT")`` R packages.
diff --git a/conf/test_use_vst.config b/conf/test_use_vst.config
index d0e416ae..41fb1d63 100644
--- a/conf/test_use_vst.config
+++ b/conf/test_use_vst.config
@@ -23,7 +23,7 @@ params {
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
     software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv'
     multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip'
-    use_vst = true
+    trafo = 'vst-force'
     genome = 'GRCm38'
     logFC_threshold = 1
 }

From aaa7d0f4d09ccf6e5eb69b61934eedda9883aa52 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 19 Jun 2023 11:33:00 +0200
Subject: [PATCH 054/124] fixed some incorrect < and layout issues

---
 CHANGELOG.md             |  2 ++
 assets/RNAseq_report.Rmd | 32 +++++++++++++++-----------------
 2 files changed, 17 insertions(+), 17 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 503fc46d..aa9f04a3 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons
 - [#205](https://github.com/qbic-pipelines/rnadeseq/pull/205) Template update
 - [#204](https://github.com/qbic-pipelines/rnadeseq/pull/204) Changed relevel path in test_relevel.config to the qbic-pipelines repo
 - [#203](https://github.com/qbic-pipelines/rnadeseq/pull/203) Switched from Dockerhub to GHCR
@@ -22,6 +23,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Fixed
 
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added correct plot titles to meanSdPlot (depending on normalization)
 - [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
 
 ## 2.1 - Wheat Fields
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index b5bbd4db..795ba95d 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -550,10 +550,11 @@ if (isProvided(params$path_relevel)) {
     }
 }
 
-# SizeFactors(cds) as indicator of library sequencing depth
+# sizeFactors(cds) as indicator of library sequencing depth
 if (params$input_type == "featurecounts") {
     sf <- sizeFactors(cds)
 } else {
+    # If tximport was used, sizeFactors doesn't work, so use workaround as described here: https://support.bioconductor.org/p/97676/
     nm <- assays(cds)[["avgTxLength"]]
     sf <- estimateSizeFactorsForMatrix(counts(cds)/nm)
 }
@@ -580,7 +581,7 @@ nsub_genes_text <- ""
 if (params$trafo == "vst-force") {
     run_rlog <- F
     trafo_long_text <- "variant stabilizing transformation (vst)"
-    nsub_genes_text <- paste0("The number of subset genes for the vst transformation is ", as.character(params$nsub_genes), ".")
+    nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
         trafo_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
         nsub_genes_text <- ""
@@ -588,7 +589,7 @@ if (params$trafo == "vst-force") {
 } else if (params$trafo == "vst") {
     run_rlog <- F
     trafo_long_text <- "variant stabilizing transformation (vst)"
-    nsub_genes_text <- paste0("The number of subset genes for the vst transformation is ", as.character(params$nsub_genes), ".")
+    nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
         run_rlog <- T
         trafo_long_text <- "regularized logarithm (rlog)"
@@ -847,7 +848,7 @@ Note: Duplicate intercept (%) is the percentage of duplicate reads for the inter
 
 ## Mapping statistics {.tabset}
 
-The read mapping statistics to the reference genome for each sample are shown below. Typical values for a good mapping lie at >80% uniquely mapped reads.
+The read mapping statistics to the reference genome for each sample are shown below. Typical values for a good mapping lie at > 80 % uniquely mapped reads.
 
 ### Percentages {-}
 
@@ -1391,8 +1392,8 @@ pval_text <- as.character(params$pval_threshold)
 ## List of differentially expressed genes
 
 The analysis of the differential gene expression was performed using `DESeq2`.
-Genes were considered differentially expressed (DE) when the adjusted p-value was lower than `r pval_text` (padj < `r pval_text`).
-Genes were further considered differentially expressed (DE) if the log2 Fold Change in expression between the two considered groups was above the threshold of `r logFC_text` in absolute terms.
+Genes were considered differentially expressed (DE) when the adjusted p-value was equal to or lower than `r pval_text` (padj <= `r pval_text`).
+Genes were further considered differentially expressed (DE) if the log2 Fold Change in expression between the two considered groups was equal to or above the threshold of `r logFC_text` in absolute terms.
 The adjusted p-value is calculated in the `DESeq2` package with the Benjamini-Hochberg method. P-value adjustment
 helps reduce the number of false postives (not real differentially expressed genes).
 
@@ -1412,7 +1413,7 @@ Additional tables for all individual contrasts are provided [here](./differentia
 
 Volcano plots display the DE genes in Log2 Fold Change values (x axis)
 against their adjusted p-value (y axis) in a form of a -log10. Only DE genes are colored red.
-Genes were considered DE with a padj < `r pval_text` (horizontal drawn through line) and in addition a Log2FC > `r logFC_text` or < -`r logFC_text` (vertical grey lines). Values above the horizontal dashed line were capped.
+Genes were considered DE with a padj <= `r pval_text` (horizontal drawn through line) and in addition a Log2FC >= `r logFC_text` or <= -`r logFC_text` (vertical grey lines). Values above the horizontal dashed line were capped.
 Here, volcano plots for all considered contrasts are shown. Hover over the dots to show the gene names.
 
 ```{r volcano_plot, out.width="100%", out.height="100%"}
@@ -1963,12 +1964,9 @@ gost_text,
 
 "\n
 ## Enriched pathways
-The plot below summarizes the pathways that were
-found significantly enriched in DE genes for each contrast (padj value < ", pval_text, ").
-Only contrasts for which an enriched pathway was found are shown.
-Both KEGG pathways and REACTOME (REAC) pathways are considered.
-Hover over the dots to reveal the pathway names.
-The table below provides more detail on all enriched pathways."))
+The plot below summarizes the pathways that were found significantly enriched in DE genes for each contrast (padj value <= ", pval_text, ").
+Only contrasts for which an enriched pathway was found are shown. Both KEGG pathways and REACTOME (REAC) pathways are considered.
+Hover over the dots to reveal the pathway names. The table below provides more detail on all enriched pathways."))
 ```
 
 ```{r gost, eval=params$pathway_analysis, out.height = "100%", out.width = "100%", warning=FALSE, message=FALSE}
@@ -2161,8 +2159,8 @@ as.character(params$revision),
 quant_tool, "` were processed with the R package `DESeq2 v",
 packageVersion("DESeq2"),
 "` [@love2014differential]. The thresholds for differentially expressed genes were set to ", 
-pval_text, "for the p value and",
-logFC_text, "for the log2 Fold Change."
+pval_text, " for the p value and ",
+logFC_text, " for the log2 Fold Change."
 ))
 
 ```{r conditional_pathway, echo=FALSE, results='asis', eval=isProvided(params$pathway_analysis)&isProvided(params$path_software_versions)}
@@ -2186,13 +2184,13 @@ if (isProvided(params$custom_gmt)){
 }
 
 cat(paste0(
-"## Differential gene expression analysis
+"\n## Pathway enrichment analysis
 
 For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2,
 " `, `AnnotationDbi v", version_annotation,
 "` and `", name_species, " v", version_annotation,
 "` were employed. ", database_string,
-"Pathways were classified as enriched for those genes with padj <= ", pval_text, "."
+"\nPathways were classified as enriched for those genes with padj <= ", pval_text, "."
 ))
 ```
 

From 6e217c88466a396654966210cca186bbb953f27a Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 19 Jun 2023 13:33:19 +0200
Subject: [PATCH 055/124] added trycatch around heatmaps

---
 CHANGELOG.md             |  2 +-
 assets/RNAseq_report.Rmd | 53 ++++++++++++++++++++++++++++------------
 2 files changed, 39 insertions(+), 16 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index d69c4496..0146b959 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
 - [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
 - [#197](https://github.com/qbic-pipelines/rnadeseq/pull/197) Added gprofiler version string to report
 - [#196](https://github.com/qbic-pipelines/rnadeseq/pull/196) Added optional `--quote` parameter
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 6d3b4a6a..ebb96ac0 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -567,7 +567,7 @@ if (params$input_type == "featurecounts") {
 write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T,  col.names = F, qmethod = c("escape", "double"))
 
 # Write cds assay table to file
-write.table(counts(cds, normalized=T), paste("differential_gene_expression/gene_counts_tables/deseq2_table.tsv", sep=""), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
+write.table(counts(cds, normalized=T), paste0("differential_gene_expression/gene_counts_tables/deseq2_table.tsv"), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
 
 # Or for rsem/salmon save folder/file names to txt file
 if (params$input_type == "rsem") {
@@ -1455,7 +1455,7 @@ log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10
 
 pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, "<br>", "Log2FC: ", formatC(log2FoldChange, digits=2)))) +
         #this goes after alpha=0.5:
-        geom_point(alpha=0.5, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold &  DE_all$logpval >= -log10(params$pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
+        geom_jitter(alpha=0.5, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold &  DE_all$logpval >= -log10(params$pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
         geom_hline(yintercept = 16, linetype= "dashed", size = 0.2, color = "grey") +
         geom_hline(yintercept = -log10(params$pval_threshold), size = 0.2, color = "grey") +
         geom_vline(xintercept = -params$logFC_threshold, size = 0.2, color = "grey") +
@@ -1835,18 +1835,41 @@ for (file in contrast_files){
                     mat$gene_name <- NULL
                     mat <- data.matrix(mat)
 
-                    # Skip the heatmaps for CI tests
+                    # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason
                     if (!isProvided(params$citest) & nrow(mat)>1){
                         dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/"))
-                        png(filename = paste("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png", sep=""), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50)
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
-                        dev.off()
-                        pdf(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf", sep=""), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
-                        dev.off()
-                        svg(paste("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg", sep=""), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
-                        pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
-                        dev.off()
+                        
+                        # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved                         
+                        tryCatch(
+                            {
+                                png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50)
+                                pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
+                                dev.off()
+                            },
+                            error=function(e) {
+                                print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png because of the following error:\n", e))
+                            }
+                        )
+                        tryCatch(
+                            {
+                                pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                                pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
+                                dev.off()
+                            },
+                            error=function(e) {
+                                print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf because of the following error:\n", e))
+                            }
+                        )
+                        tryCatch(
+                            {
+                                svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+                                pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T)
+                                dev.off()
+                            },
+                            error=function(e) {
+                                print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg because of the following error:\n", e))
+                            }
+                        )
                     }
 
                     # Plotting pathway view only for kegg pathways
@@ -1922,13 +1945,13 @@ if (isProvided(params$path_genelist)){
     mat <- data.matrix(mat)
 
     if (nrow(mat)>1){
-        png(filename = paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png", sep=""), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100)
+        png(filename = paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100)
         pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
-        pdf(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf", sep=""), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005)
+        pdf(paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf"), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005)
         pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
-        svg(paste("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg", sep=""), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
+        svg(paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg"), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005)
         pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T)
         dev.off()
     }

From df0fb8eb303427afcc3014759a81a27f5a9f8fd1 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 20 Jun 2023 09:10:29 +0200
Subject: [PATCH 056/124] removed pipe bind

---
 assets/RNAseq_report.Rmd | 5 +----
 1 file changed, 1 insertion(+), 4 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index ebb96ac0..af81dbaf 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -56,11 +56,8 @@ params:
 ---
 
 ```{r init, include=F}
-# The following envvar is necessary to enable <= and >= comparisons
-Sys.setenv("_R_USE_PIPEBIND_" = TRUE)
-
 # set a seed in case some background calculations are random-based
-    set.seed(10)
+set.seed(10)
 
 # svg path for the little svg format icon displayed in the interactive plots for svg export; based on https://www.svgviewer.dev/:
 svg_icon_path <- 'M228 319.5H12C5.64873 319.5 0.5 314.351 0.5 308V12C0.5 5.64873 5.64873 0.5 12 0.5H159.175C159.571 0.5 159.952 0.656748 160.233 0.936021L239.057 79.2681C239.341 79.5497 239.5 79.9326 239.5 80.3321V308C239.5 314.351 234.351 319.5 228 319.5Z'

From 3610ddf0bde6052247f068713835fb1df51370e7 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 09:30:50 +0200
Subject: [PATCH 057/124] Update assets/RNAseq_report.Rmd

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 assets/RNAseq_report.Rmd | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index af81dbaf..59b81324 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -583,7 +583,7 @@ trafo_override_text <- ""
 nsub_genes_text <- ""
 if (params$trafo == "vst-force") {
     run_rlog <- F
-    trafo_long_text <- "variant stabilizing transformation (vst)"
+    trafo_long_text <- "variance stabilizing transformation (vst)"
     nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
         trafo_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")

From 3b1e4ca817230d517214a97f7abb727a66b5ab52 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 09:30:59 +0200
Subject: [PATCH 058/124] Update assets/RNAseq_report.Rmd

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 assets/RNAseq_report.Rmd | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 59b81324..d355ec3c 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -591,7 +591,7 @@ if (params$trafo == "vst-force") {
     }
 } else if (params$trafo == "vst") {
     run_rlog <- F
-    trafo_long_text <- "variant stabilizing transformation (vst)"
+    trafo_long_text <- "variance stabilizing transformation (vst)"
     nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
         run_rlog <- T

From ce4f34971202e4b69d97dfeb262320e93bd99ff7 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 10:05:20 +0200
Subject: [PATCH 059/124] Update assets/RNAseq_report.Rmd

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 assets/RNAseq_report.Rmd | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index d355ec3c..f6b9227e 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -2214,7 +2214,7 @@ cat(paste0(
 For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2,
 " `, `AnnotationDbi v", version_annotation,
 "` and `", name_species, " v", version_annotation,
-"` were employed. ", database_string, ".\n",
+"` were used. ", database_string, ".\n",
 "Pathways were classified as enriched for those genes with padj <= ", pval_text, "."
 ))
 ```

From c326b6b47e5b88f6c39896c6775f3681aebc9697 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 10:08:13 +0200
Subject: [PATCH 060/124] Update docs/usage.md

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index d7078cf1..eabaa4d7 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -307,7 +307,7 @@ Integer indicating how many genes in a pathway must be differentially expressed
 
 ### `--trafo`
 
-Set to `vst`, `vst-force` or `rlog` (default) to determine which transformation is applied to the data. If `vst`, the pipeline can still use rlog instead if the input data are found to have different size factor variances (>=0.05) which will be stated in the report. If you want to use the vst method regardless of data, use `vst-force`. Generally speaking, vst is faster but rlog is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
+Set  this parameter to either `vst`, `vst-force` or `rlog` (default) to control which transformation should be applied to the data. If the input data has too large size factor variances (>=0.05) the pipeline will override a user-specified `vst` transformation to use `rlog` instead. If this is enforced it will be stated in the report. If you still prefer to use the `vst` method regardless of the variances in the data, please use the `vst-force` parameter. While the `vst` transformation is much faster, the `rlog` is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
 
 ### `--vst_genes_number`
 

From e2abe68151316b2ec02f37f7adf079ab2da9c01f Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 10:08:52 +0200
Subject: [PATCH 061/124] Update docs/usage.md

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index eabaa4d7..dffd8367 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -311,7 +311,7 @@ Set  this parameter to either `vst`, `vst-force` or `rlog` (default) to control
 
 ### `--vst_genes_number`
 
-This is ignored if `--trafo` is set to `rlog`. If using the vst transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small rnaseq data. The default `vst` function for varianceStabilizingTransformation in DESeq2 is 1000, which triggers an error with small datasets. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
+This is ignored if `--trafo` is set to `rlog`. If using the `vst` transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small RNA-Seq data. The default number of genes for applying the `vst` function for varianceStabilizingTransformation in DESeq2 is 1000. For smaller datasets there will be an error. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
 
 ### `--round_DE`
 

From 4470d2ae5378207938a8c475038fb1c41f4afd7d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 20 Jun 2023 10:32:44 +0200
Subject: [PATCH 062/124] changed some things for the review

---
 CHANGELOG.md             |  2 +-
 assets/RNAseq_report.Rmd | 59 ++++++++++++++++++++--------------------
 bin/Execute_report.R     |  4 +--
 docs/usage.md            |  4 +--
 modules/local/report.nf  |  3 +-
 nextflow.config          |  2 +-
 nextflow_schema.json     |  2 +-
 7 files changed, 38 insertions(+), 38 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 0146b959..2985a922 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -14,7 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
-- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_table.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
 - [#205](https://github.com/qbic-pipelines/rnadeseq/pull/205) Template update
 - [#204](https://github.com/qbic-pipelines/rnadeseq/pull/204) Changed relevel path in test_relevel.config to the qbic-pipelines repo
 - [#203](https://github.com/qbic-pipelines/rnadeseq/pull/203) Switched from Dockerhub to GHCR
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index af81dbaf..09f8e9d0 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -28,7 +28,7 @@ params:
     path_relevel: ''
     batch_effect: ''
     logFC_threshold: ''
-    pval_threshold: ''
+    adj_pval_threshold: ''
     trafo: ''
     nsub_genes: ''
     round_DE: ''
@@ -564,7 +564,7 @@ if (params$input_type == "featurecounts") {
 write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T,  col.names = F, qmethod = c("escape", "double"))
 
 # Write cds assay table to file
-write.table(counts(cds, normalized=T), paste0("differential_gene_expression/gene_counts_tables/deseq2_table.tsv"), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
+write.table(counts(cds, normalized=T), paste0("differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv"), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
 
 # Or for rsem/salmon save folder/file names to txt file
 if (params$input_type == "rsem") {
@@ -705,18 +705,18 @@ for (i in resultsNames(cds)[-1]) {
     # rename the levels of the bins using the middle point
     levels(bins) <- paste0("~",round(.5*qs[-1] + .5*qs[-length(qs)]))
 
-    # calculate the ratio of p values less than .01 for each bin
+    # calculate the ratio of p-values less than .01 for each bin
     ratios <- tapply(res$pvalue, bins, function(p) mean(p < .01, na.rm=TRUE ))
 
     # plot these ratios
     pdf(paste("differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_",i,".pdf",sep=""))
-    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p values")
+    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p-values")
     dev.off()
     png(paste("differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_",i,".png",sep=""))
-    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p values")
+    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p-values")
     dev.off()
     svg(paste("differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_",i,".svg",sep=""))
-    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p values")
+    barplot(ratios, xlab="mean normalized count", ylab="ratio of small p-values")
     dev.off()
 
     # plot number of rejections
@@ -750,21 +750,21 @@ for (i in resultsNames(cds)[-1]) {
     colori <- c('do not pass'="khaki", 'pass'="powderblue")
     pdf(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".pdf",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
-            col = colori, space = 0, main = "", xlab="p value",ylab="frequency")
+            col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
         adj = c(0.5,1.7), xpd=NA)
     legend("topleft", fill=rev(colori), legend=rev(names(colori)))
     dev.off()
     png(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".png",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
-            col = colori, space = 0, main = "", xlab="p value",ylab="frequency")
+            col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
         adj = c(0.5,1.7), xpd=NA)
     legend("topleft", fill=rev(colori), legend=rev(names(colori)))
     dev.off()
     svg(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".svg",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
-            col = colori, space = 0, main = "", xlab="p value",ylab="frequency")
+            col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
         adj = c(0.5,1.7), xpd=NA)
     legend("topleft", fill=rev(colori), legend=rev(names(colori)))
@@ -964,7 +964,7 @@ cat(paste0("***
 
 The raw count table and normalized count tables are available [here](./differential_gene_expression/gene_counts_tables).
 The differential expression analysis is performed using the raw gene count table.
-For PCA analysis and heatmap plotting, the `r trafo_long_text` normalized gene counts were employed. 
+For PCA analysis and heatmap plotting, the `r trafo_long_text` normalized gene counts were used. 
 `r trafo_override_text`
 `r nsub_genes_text`
 
@@ -1099,6 +1099,7 @@ After unzipping the results ZIP file, the differential expression results can be
     `r rsem_salmon_string`
 - `gene_counts_tables/`
     `r featurecounts_string`
+    - `deseq2_library_scaled_gene_counts.tsv`: gene counts after size factor scaling.
     `r genecounts_string`
     - `sizeFactor_libraries.tsv`: size factors for each sample.
 - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast.
@@ -1164,7 +1165,7 @@ if (isProvided(params$path_contrast_matrix)){
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1224,7 +1225,7 @@ if (isProvided(params$path_contrast_list)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1266,7 +1267,7 @@ if (isProvided(params$path_contrast_pairs)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
 
         # Round table before writing
@@ -1306,7 +1307,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE)) {
@@ -1339,8 +1340,8 @@ logFC = DE_genes_df[,logFC_cols,drop=F]
 padj = DE_genes_df[,padj_cols,drop=F]
 padj[is.na(padj)] <- 1
 
-# Convert to binary (1/0) matrix if padj <= params$pval_threshold or not, respectively
-padj_bin = data.matrix(ifelse(padj <= params$pval_threshold, 1, 0))
+# Convert to binary (1/0) matrix if padj <= params$adj_pval_threshold or not, respectively
+padj_bin = data.matrix(ifelse(padj <= params$adj_pval_threshold, 1, 0))
 
 # Convert to binary (1/0) matrix if logFC is bigger/equal or smaller/equal than threshold or not, respectively
 logFC_bin = data.matrix(ifelse(abs(logFC) >= params$logFC_threshold, 1, 0))
@@ -1389,7 +1390,7 @@ for (i in 1:length(contrast_names)){
 
 ```{r load_config, bootstrap.show.message=FALSE, echo=FALSE, message=FALSE, warning=FALSE}
 logFC_text <- as.character(params$logFC_threshold)
-pval_text <- as.character(params$pval_threshold)
+pval_text <- as.character(params$adj_pval_threshold)
 ```
 
 ## List of differentially expressed genes
@@ -1452,9 +1453,9 @@ log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10
 
 pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, "<br>", "Log2FC: ", formatC(log2FoldChange, digits=2)))) +
         #this goes after alpha=0.5:
-        geom_jitter(alpha=0.5, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold &  DE_all$logpval >= -log10(params$pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
+        geom_jitter(alpha=0.5, width = 0.2, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold &  DE_all$logpval >= -log10(params$adj_pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) +
         geom_hline(yintercept = 16, linetype= "dashed", size = 0.2, color = "grey") +
-        geom_hline(yintercept = -log10(params$pval_threshold), size = 0.2, color = "grey") +
+        geom_hline(yintercept = -log10(params$adj_pval_threshold), size = 0.2, color = "grey") +
         geom_vline(xintercept = -params$logFC_threshold, size = 0.2, color = "grey") +
         geom_vline(xintercept = params$logFC_threshold, size = 0.2, color = "grey") +
         scale_color_manual(name = "", values=c("red", "black")) +
@@ -1465,8 +1466,8 @@ pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_
         scale_x_continuous(breaks = seq(-200, 200, ifelse(log2FoldChange_quotient<3, 1, 5)))
 
 pg <- ggplotly(pg, tooltip = "text")
-pg[['x']][['layout']][['annotations']][[2]][['x']] <- -params$pval_threshold #-0.05 TODO: what do these two lines do?
-pg[['x']][['layout']][['annotations']][[1]][['y']] <- -params$pval_threshold
+pg[['x']][['layout']][['annotations']][[2]][['x']] <- -params$adj_pval_threshold #-0.05 TODO: what do these two lines do?
+pg[['x']][['layout']][['annotations']][[1]][['y']] <- -params$adj_pval_threshold
 
 #add export button to plot
 config(pg, modeBarButtonsToAdd = list(svg_exp)) %>% layout(margin = list(l=80, b = ifelse(length(allgenes_files)>2, 150, 100)), width=700, height = 500*length(allgenes_files))
@@ -1692,7 +1693,7 @@ for (file in contrast_files){
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
-            user_threshold=params$pval_threshold,
+            user_threshold=params$adj_pval_threshold,
             custom_bg=custom_background,
             domain_scope="custom_annotated"
         )
@@ -1708,7 +1709,7 @@ for (file in contrast_files){
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
-            user_threshold=params$pval_threshold,
+            user_threshold=params$adj_pval_threshold,
             domain_scope="annotated"
         )
         pathway_gostres_nobg <- gostres_nobg$result
@@ -1722,7 +1723,7 @@ for (file in contrast_files){
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
-            user_threshold=params$pval_threshold,
+            user_threshold=params$adj_pval_threshold,
             domain_scope="annotated"
         )
     }
@@ -2026,7 +2027,7 @@ if (length(q_list) > 0) {
                     significant=T,
                     correction_method="fdr",
                     sources=datasources,
-                    user_threshold=params$pval_threshold,
+                    user_threshold=params$adj_pval_threshold,
                     custom_bg=custom_background,
                     domain_scope="custom_annotated"
         )
@@ -2036,7 +2037,7 @@ if (length(q_list) > 0) {
                     significant=T,
                     correction_method="fdr",
                     sources=datasources,
-                    user_threshold=params$pval_threshold,
+                    user_threshold=params$adj_pval_threshold,
                     domain_scope="annotated"
         )
     }
@@ -2044,7 +2045,7 @@ if (length(q_list) > 0) {
     path_enrich <- as.data.frame(gostres$result)
     if (nrow(path_enrich) > 0){
         pg2 <- gostplot(gostres, capped=T, interactive=T)
-        pg2[['x']][['layout']][['annotations']][[1]][['x']] <- -params$pval_threshold
+        pg2[['x']][['layout']][['annotations']][[1]][['x']] <- -params$adj_pval_threshold
 
         #limit gostplot y maximum
         for (counter in c(1:length(contrast_files))) {
@@ -2185,7 +2186,7 @@ as.character(params$revision),
 quant_tool, "` were processed with the R package `DESeq2 v",
 packageVersion("DESeq2"),
 "` [@love2014differential]. The thresholds for differentially expressed genes were set to ", 
-pval_text, " for the p value and ",
+pval_text, " for the p-value and ",
 logFC_text, " for the log2 Fold Change."
 ))
 
@@ -2215,7 +2216,7 @@ For pathway analysis, the R packages `gprofiler2 v", version_gprofiler2,
 " `, `AnnotationDbi v", version_annotation,
 "` and `", name_species, " v", version_annotation,
 "` were employed. ", database_string, ".\n",
-"Pathways were classified as enriched for those genes with padj <= ", pval_text, "."
+"Pathways were classified as enriched for those genes with an adjusted p-value <= ", pval_text, "."
 ))
 ```
 
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index a3573582..ea2c7c3a 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -20,7 +20,7 @@ option_list = list(
     make_option(c("-e", "--relevel"), type="character", default=NULL, help="Path to factor relevel file", metavar="character"),
     make_option(c("-b", "--batch_effect"), action="store_true", default=FALSE, help="Batch effect correction."),
     make_option(c("-f", "--logFC_threshold"), type="double", default=NULL, help="Log Fold Change threshold to consider a gene DE."),
-    make_option("--pval_threshold", type="double", default=0.05, help="p value threshold to consider a gene DE."),
+    make_option("--adj_pval_threshold", type="double", default=0.05, help="p value threshold to consider a gene DE."),
     make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
 
     #make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
@@ -70,7 +70,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 path_relevel = opt$relevel,
                                 batch_effect = opt$batch_effect,
                                 logFC_threshold = opt$logFC_threshold,
-                                pval_threshold = opt$pval_threshold,
+                                adj_pval_threshold = opt$adj_pval_threshold,
                                 trafo = opt$trafo,
                                 nsub_genes = opt$nsub_genes,
                                 round_DE = opt$round_DE,
diff --git a/docs/usage.md b/docs/usage.md
index d7078cf1..3d09558b 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -26,7 +26,7 @@
   - [`--contrast_pairs`](#--contrast_pairs)
 - [Optional arguments](#optional-arguments)
   - [`--logFC_threshold`](#--logFC_threshold)
-  - [`--pval_threshold`](#--pval_threshold)
+  - [`--adj_pval_threshold`](#--adj_pval_threshold)
   - [`--genelist`](#--genelist)
   - [`--batch_effect`](#--batch_effect)
   - [`--min_DEG_pathway`](#--min_deg_pathway)
@@ -277,7 +277,7 @@ interaction_effect condition_treatment_treated_vs_control  condition_genotype_KO
 
 Threshold (int) to apply to Log 2 Fold Change to consider a gene as differentially expressed. There is no threshold applied by default to Log2 Fold Change.
 
-### `--pval_threshold`
+### `--adj_pval_threshold`
 
 P value (float) to consider a gene as differentially expressed. The default value is 0.05.
 
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 556ebad5..c68b1e4a 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -32,7 +32,6 @@ process REPORT {
     def genelist_opt = genelist.name != 'NO_FILE' ? "--genelist $genelist" : ''
     def relevel_opt = relevel.name != 'NO_FILE2' ? "--relevel $relevel" : ''
     def batch_effect_opt = params.batch_effect ? "--batch_effect TRUE" : ''
-//    def rlog_opt = params.use_vst ? '--rlog FALSE' : ''
     def round_DE_opt = params.round_DE ? "--round_DE $params.round_DE" : ''
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
@@ -66,7 +65,7 @@ process REPORT {
         $relevel_opt \
         $batch_effect_opt \
         --logFC_threshold $params.logFC_threshold \
-        --pval_threshold $params.pval_threshold \
+        --adj_pval_threshold $params.adj_pval_threshold \
         --trafo $params.trafo \
         --nsub_genes $params.vst_genes_number \
         $round_DE_opt \
diff --git a/nextflow.config b/nextflow.config
index 034f4d64..d84fc80c 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -25,7 +25,7 @@ params {
     relevel = 'NO_FILE2'
     batch_effect = null
     logFC_threshold = 0
-    pval_threshold = 0.05
+    adj_pval_threshold = 0.05
     trafo = 'rlog'
     vst_genes_number = 1000
     round_DE = null
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 93d0fb07..df1712ce 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -75,7 +75,7 @@
                     "default": 0,
                     "description": "Threshold (int) to apply to Log 2 Fold Change to consider a gene as differentially expressed."
                 },
-                "pval_threshold": {
+                "adj_pval_threshold": {
                     "type": "number",
                     "default": 0.05,
                     "description": "p value (float) to consider a gene as differentially expressed. The default value is 0.05."

From 06382e0d6a87c77004824b7932235c8c0e893f9f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 20 Jun 2023 10:53:01 +0200
Subject: [PATCH 063/124] prettier

---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index 4d96a3a6..ed6215da 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -307,7 +307,7 @@ Integer indicating how many genes in a pathway must be differentially expressed
 
 ### `--trafo`
 
-Set  this parameter to either `vst`, `vst-force` or `rlog` (default) to control which transformation should be applied to the data. If the input data has too large size factor variances (>=0.05) the pipeline will override a user-specified `vst` transformation to use `rlog` instead. If this is enforced it will be stated in the report. If you still prefer to use the `vst` method regardless of the variances in the data, please use the `vst-force` parameter. While the `vst` transformation is much faster, the `rlog` is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
+Set this parameter to either `vst`, `vst-force` or `rlog` (default) to control which transformation should be applied to the data. If the input data has too large size factor variances (>=0.05) the pipeline will override a user-specified `vst` transformation to use `rlog` instead. If this is enforced it will be stated in the report. If you still prefer to use the `vst` method regardless of the variances in the data, please use the `vst-force` parameter. While the `vst` transformation is much faster, the `rlog` is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
 
 ### `--vst_genes_number`
 

From 9e0666a219416a2449614839e733f19e8b849248 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 20 Jun 2023 11:09:15 +0200
Subject: [PATCH 064/124] renamed trafo to norm_method, updated test configs

---
 CHANGELOG.md                   |  2 +-
 assets/RNAseq_report.Rmd       | 38 +++++++++++++++++-----------------
 bin/Execute_report.R           |  4 ++--
 conf/test_use_vst.config       |  2 +-
 docs/usage.md                  |  6 +++---
 modules/local/report.nf        |  2 +-
 nextflow.config                |  2 +-
 nextflow_schema.json           |  2 +-
 tests/test.yml                 |  2 +-
 tests/test_batcheffect.yml     |  2 +-
 tests/test_contrast_list.yml   |  2 +-
 tests/test_contrast_matrix.yml |  2 +-
 tests/test_custom_gmt.yml      |  2 +-
 tests/test_full.yml            |  2 +-
 tests/test_no_multiqc.yml      |  2 +-
 tests/test_relevel.yml         |  2 +-
 tests/test_star_salmon.yml     |  2 +-
 tests/test_use_vst.yml         |  2 +-
 18 files changed, 39 insertions(+), 39 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 2985a922..60c94179 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -14,7 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
-- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_table.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
+- [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_library_scaled_gene_counts.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
 - [#205](https://github.com/qbic-pipelines/rnadeseq/pull/205) Template update
 - [#204](https://github.com/qbic-pipelines/rnadeseq/pull/204) Changed relevel path in test_relevel.config to the qbic-pipelines repo
 - [#203](https://github.com/qbic-pipelines/rnadeseq/pull/203) Switched from Dockerhub to GHCR
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 02c0f4b7..787674e3 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -29,7 +29,7 @@ params:
     batch_effect: ''
     logFC_threshold: ''
     adj_pval_threshold: ''
-    trafo: ''
+    norm_method: ''
     nsub_genes: ''
     round_DE: ''
 
@@ -578,29 +578,29 @@ if (params$input_type == "rsem") {
 vst_cutoff <- 0.05
 var_sizeFactors <- var(sf)
 run_rlog <- T
-trafo_long_text <- "regularized logarithm (rlog)"
-trafo_override_text <- ""
+norm_method_long_text <- "regularized logarithm (rlog)"
+norm_method_override_text <- ""
 nsub_genes_text <- ""
-if (params$trafo == "vst-force") {
+if (params$norm_method == "vst-force") {
     run_rlog <- F
-    trafo_long_text <- "variance stabilizing transformation (vst)"
+    norm_method_long_text <- "variance stabilizing transformation (vst)"
     nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
-        trafo_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
+        norm_method_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
         nsub_genes_text <- ""
     }
-} else if (params$trafo == "vst") {
+} else if (params$norm_method == "vst") {
     run_rlog <- F
-    trafo_long_text <- "variance stabilizing transformation (vst)"
+    norm_method_long_text <- "variance stabilizing transformation (vst)"
     nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
         run_rlog <- T
-        trafo_long_text <- "regularized logarithm (rlog)"
-        trafo_override_text <- paste0(" This was done as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
+        norm_method_long_text <- "regularized logarithm (rlog)"
+        norm_method_override_text <- paste0(" This was done as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
         nsub_genes_text <- ""
     }
 }
-trafo_text <- ifelse(run_rlog, "rlog", "vst")
+norm_method_text <- ifelse(run_rlog, "rlog", "vst")
 
 if (run_rlog){
 
@@ -667,21 +667,21 @@ pdf("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_tra
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", norm_method_text, "transformed count data"))
 dev.off()
 notAllZero <- (rowSums(counts(cds))>0)
 png("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png")
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", norm_method_text, "transformed count data"))
 dev.off()
 notAllZero <- (rowSums(counts(cds))>0)
 svg("differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg")
 par(oma=c(3,3,3,3))
 par(mfrow = c(1, 3))
 meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab  = "sd raw count data")
-meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", trafo_text, "transformed count data"))
+meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab  = paste("sd", norm_method_text, "transformed count data"))
 dev.off()
 
 # Further diagnostics plots
@@ -964,13 +964,13 @@ cat(paste0("***
 
 The raw count table and normalized count tables are available [here](./differential_gene_expression/gene_counts_tables).
 The differential expression analysis is performed using the raw gene count table.
-For PCA analysis and heatmap plotting, the `r trafo_long_text` normalized gene counts were used. 
-`r trafo_override_text`
+For PCA analysis and heatmap plotting, the `r norm_method_long_text` normalized gene counts were used. 
+`r norm_method_override_text`
 `r nsub_genes_text`
 
 ## Principal component analysis (PCA)
 
-A PCA plot of the `r trafo_text` normalized gene expression visualizes the clustering of samples according to their gene expression.
+A PCA plot of the `r norm_method_text` normalized gene expression visualizes the clustering of samples according to their gene expression.
 In this way, the overall effect of experimental conditions and sample grouping, and any batch effects are visualized.
 The original plot can be downloaded [here](./differential_gene_expression/plots/PCA_plot.pdf).
 
@@ -1105,8 +1105,8 @@ After unzipping the results ZIP file, the differential expression results can be
 - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast.
 - `final_gene_table/final_gene_list_DESeq2.tsv`: table containing a list of all genes considered in the analysis. Here a summary of the log2 Fold Change and p-adjusted values for all contrasts is displayed. Additionally, the column **filter** shows if this gene was differentially expressed (DE) in any of the contrasts, or not (not_DE). The column **contrast_vector** contains for each contrast considered in the analysis a 1 if the gene was differentially expressed for this contrast or a 0 if it was not.
 - `plots/`:
-    - `Heatmaps_of_distances.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r trafo_text` normalized gene counts are considered.
-    - `PCA_plot.pdf`: PCA of the `r trafo_text` normalized gene counts.
+    - `Heatmaps_of_distances.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered.
+    - `PCA_plot.pdf`: PCA of the `r norm_method_text` normalized gene counts.
     - `boxplots_example_genes/`: boxplots of the normalized gene counts for each of the sample groups for example genes.
     - `boxplots_requested_genes/`: boxplots of the normalized gene counts for each of the sample groups for the list of requested genes.
     - `further_diagnostics_plots/`: plots for diagnostics of the differential gene analysis procedure.
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index ea2c7c3a..5d413a2c 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -24,7 +24,7 @@ option_list = list(
     make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
 
     #make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
-    make_option(c("-z", "--trafo"), type="character", default=NULL, help="Which transformation(s) to use."),
+    make_option(c("-z", "--norm_method"), type="character", default=NULL, help="Which transformation(s) to use."),
     make_option(c("-n", "--nsub_genes"), type="integer", default=NULL, help="Subset number of genes for vst."),
 
     make_option(c("-a", "--pathway_analysis"), action="store_true", default=FALSE, help="Whether to run pathway analysis."),
@@ -71,7 +71,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp
                                 batch_effect = opt$batch_effect,
                                 logFC_threshold = opt$logFC_threshold,
                                 adj_pval_threshold = opt$adj_pval_threshold,
-                                trafo = opt$trafo,
+                                norm_method = opt$norm_method,
                                 nsub_genes = opt$nsub_genes,
                                 round_DE = opt$round_DE,
 
diff --git a/conf/test_use_vst.config b/conf/test_use_vst.config
index 41fb1d63..9e81793a 100644
--- a/conf/test_use_vst.config
+++ b/conf/test_use_vst.config
@@ -23,7 +23,7 @@ params {
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
     software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv'
     multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip'
-    trafo = 'vst-force'
+    norm_method = 'vst-force'
     genome = 'GRCm38'
     logFC_threshold = 1
 }
diff --git a/docs/usage.md b/docs/usage.md
index ed6215da..9b70d8e3 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -30,7 +30,7 @@
   - [`--genelist`](#--genelist)
   - [`--batch_effect`](#--batch_effect)
   - [`--min_DEG_pathway`](#--min_deg_pathway)
-  - [`--trafo`](#--trafo)
+  - [`--norm_method`](#--norm_method)
   - [`--vst_genes_number`](#--vst_genes_number)
   - [`--round_DE`](#--round_DE)
   - [`--run_pathway_analysis`](#--run_pathway_analysis)
@@ -305,13 +305,13 @@ For more information, please check the [DESeq2 vignette](http://bioconductor.org
 
 Integer indicating how many genes in a pathway must be differentially expressed to be considered as enriched, and report these pathways in tables and the final report. The default value is 1.
 
-### `--trafo`
+### `--norm_method`
 
 Set this parameter to either `vst`, `vst-force` or `rlog` (default) to control which transformation should be applied to the data. If the input data has too large size factor variances (>=0.05) the pipeline will override a user-specified `vst` transformation to use `rlog` instead. If this is enforced it will be stated in the report. If you still prefer to use the `vst` method regardless of the variances in the data, please use the `vst-force` parameter. While the `vst` transformation is much faster, the `rlog` is better suited to account for different sequencing depths between samples. Check here for more information on [count data transformations](https://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#count-data-transformations).
 
 ### `--vst_genes_number`
 
-This is ignored if `--trafo` is set to `rlog`. If using the `vst` transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small RNA-Seq data. The default number of genes for applying the `vst` function for varianceStabilizingTransformation in DESeq2 is 1000. For smaller datasets there will be an error. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
+This is ignored if `--norm_method` is set to `rlog`. If using the `vst` transformation, consider using this parameter for small datasets and low numbers of genes, e.g. with small RNA-Seq data. The default number of genes for applying the `vst` function for varianceStabilizingTransformation in DESeq2 is 1000. For smaller datasets there will be an error. The solution is to reduce the number of genes to sample for the transformation ( < 1000 ). More information/solution here: [DESeq2 vst function error](https://www.biostars.org/p/456209/).
 
 ### `--round_DE`
 
diff --git a/modules/local/report.nf b/modules/local/report.nf
index c68b1e4a..8a4e235f 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -66,7 +66,7 @@ process REPORT {
         $batch_effect_opt \
         --logFC_threshold $params.logFC_threshold \
         --adj_pval_threshold $params.adj_pval_threshold \
-        --trafo $params.trafo \
+        --norm_method $params.norm_method \
         --nsub_genes $params.vst_genes_number \
         $round_DE_opt \
         $pathway_opt \
diff --git a/nextflow.config b/nextflow.config
index d84fc80c..f4881c88 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -26,7 +26,7 @@ params {
     batch_effect = null
     logFC_threshold = 0
     adj_pval_threshold = 0.05
-    trafo = 'rlog'
+    norm_method = 'rlog'
     vst_genes_number = 1000
     round_DE = null
 
diff --git a/nextflow_schema.json b/nextflow_schema.json
index df1712ce..a7ede838 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -80,7 +80,7 @@
                     "default": 0.05,
                     "description": "p value (float) to consider a gene as differentially expressed. The default value is 0.05."
                 },
-                "trafo": {
+                "norm_method": {
                     "type": "string",
                     "default": "rlog",
                     "enum": ["rlog", "vst", "vst-force"],
diff --git a/tests/test.yml b/tests/test.yml
index 4b9b6105..2adebac0 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml
index 216f3301..72150e5b 100644
--- a/tests/test_batcheffect.yml
+++ b/tests/test_batcheffect.yml
@@ -9,7 +9,7 @@
       # md5sum: 2738119f899ecd154cd5cfc40e1e24e4
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       # md5sum: b566de65d7b6e55aad648f74c439fcea
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_contrast_list.yml b/tests/test_contrast_list.yml
index a71ea0be..f3aebcd6 100644
--- a/tests/test_contrast_list.yml
+++ b/tests/test_contrast_list.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 92323960e7328056d35bbc6c668a2d06
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_contrast_matrix.yml b/tests/test_contrast_matrix.yml
index 3763518b..05d5a02b 100644
--- a/tests/test_contrast_matrix.yml
+++ b/tests/test_contrast_matrix.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 7950e3e0a5084e431134aa943884a488
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml
index 5a9dec79..071c297b 100644
--- a/tests/test_custom_gmt.yml
+++ b/tests/test_custom_gmt.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_full.yml b/tests/test_full.yml
index 77abaeee..21be0ef4 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_no_multiqc.yml b/tests/test_no_multiqc.yml
index b67fd5ac..9e86ef4d 100644
--- a/tests/test_no_multiqc.yml
+++ b/tests/test_no_multiqc.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml
index b9cccb46..92e927d6 100644
--- a/tests/test_relevel.yml
+++ b/tests/test_relevel.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index bc2db1bb..3a8dc898 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -13,7 +13,7 @@
       md5sum: fcfd60285199bb7adfe0ec001f95f86f
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: e493ccfc9f15d05049776a424b1464f1
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL10_ENSG00000136634.pdf
diff --git a/tests/test_use_vst.yml b/tests/test_use_vst.yml
index 20fd23a4..eb8a731a 100644
--- a/tests/test_use_vst.yml
+++ b/tests/test_use_vst.yml
@@ -9,7 +9,7 @@
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 81d3437334849b3b4de4de6729a3d54d
-    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_table.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv

From 5fbb30fb03e3c746158fea67d603162762bb6f93 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 20 Jun 2023 13:23:27 +0200
Subject: [PATCH 065/124] updated docs

---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index 9b70d8e3..a2e16139 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -279,7 +279,7 @@ Threshold (int) to apply to Log 2 Fold Change to consider a gene as differential
 
 ### `--adj_pval_threshold`
 
-P value (float) to consider a gene as differentially expressed. The default value is 0.05.
+Adjusted p-value (float) to consider a gene as differentially expressed. The default value is 0.05.
 
 ### `--genelist`
 

From adef6aa7445fd8376b7415f95af9b224ea089b4f Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 13:36:18 +0200
Subject: [PATCH 066/124] Update bin/Execute_report.R

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 bin/Execute_report.R | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index 5d413a2c..69cc1a42 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -20,7 +20,7 @@ option_list = list(
     make_option(c("-e", "--relevel"), type="character", default=NULL, help="Path to factor relevel file", metavar="character"),
     make_option(c("-b", "--batch_effect"), action="store_true", default=FALSE, help="Batch effect correction."),
     make_option(c("-f", "--logFC_threshold"), type="double", default=NULL, help="Log Fold Change threshold to consider a gene DE."),
-    make_option("--adj_pval_threshold", type="double", default=0.05, help="p value threshold to consider a gene DE."),
+    make_option("--adj_pval_threshold", type="double", default=0.05, help="adjusted p value threshold to consider a gene DE."),
     make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
 
     #make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),

From 646ffa4678cf7ca7c4e576b4a560843ba583b1b2 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Tue, 20 Jun 2023 13:36:48 +0200
Subject: [PATCH 067/124] Update bin/Execute_report.R

Co-authored-by: SusiJo <43847534+SusiJo@users.noreply.github.com>
---
 bin/Execute_report.R | 1 -
 1 file changed, 1 deletion(-)

diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index 69cc1a42..c0463125 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -23,7 +23,6 @@ option_list = list(
     make_option("--adj_pval_threshold", type="double", default=0.05, help="adjusted p value threshold to consider a gene DE."),
     make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
 
-    #make_option(c("-z", "--rlog"), action="store_true", default=TRUE, help="Use rlog instead of vst normalization."),
     make_option(c("-z", "--norm_method"), type="character", default=NULL, help="Which transformation(s) to use."),
     make_option(c("-n", "--nsub_genes"), type="integer", default=NULL, help="Subset number of genes for vst."),
 

From 339f67257d863fe36926fa382894034e55ab8b53 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 30 Jun 2023 08:45:18 +0200
Subject: [PATCH 068/124] fixed releve bug, changelog is already updated

---
 CHANGELOG.md             | 1 +
 assets/RNAseq_report.Rmd | 8 ++++----
 2 files changed, 5 insertions(+), 4 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 60c94179..93201a2e 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -23,6 +23,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Fixed
 
+- [#208](https://github.com/qbic-pipelines/rnadeseq/pull/208) Fixed relevel bug, the function should now finally work!
 - [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added correct plot titles to meanSdPlot (depending on normalization)
 - [#195](https://github.com/qbic-pipelines/rnadeseq/pull/195) Fixed section error in report
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 787674e3..1c73c05f 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -449,7 +449,6 @@ DT::datatable(df_QA_neat, options = list(scrollX  = "100%", scrollY = "400px", p
 # Run DESeq function
 if (params$input_type == "featurecounts") {
     cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]]))))
-    cds <- DESeq(cds,  parallel = FALSE)
 } else if (params$input_type %in% c("rsem", "salmon")) {
     ## Create a dataframe which consists of both the gene id and the transcript name
     gtf <- rtracklayer::import(params$path_gtf)
@@ -504,7 +503,6 @@ if (params$input_type == "featurecounts") {
 
         #dds from SummarizedExperiment <se>, then run DESeq
         cds <- DESeqDataSet(se, design = as.formula(eval(parse(text=as.character(design[[1]])))))
-        cds <- DESeq(cds)
 
 # Load salmon count files
     } else if (params$input_type == "salmon") {
@@ -537,13 +535,12 @@ if (params$input_type == "featurecounts") {
         coldata$combfactor <- metadata$combfactor
         rownames(coldata) <- qbicCodes
         cds <- DESeqDataSetFromTximport(txi=txi.salmon, colData =coldata, design = eval(parse(text=as.character(design[[1]]))))
-        cds <- DESeq(cds)
     }
 } else {
     stop(paste0("Invalid input type: ", params$input_type, "! Input type must be one of: featurecounts, rsem, salmon!"))
 }
 
-#Apply relevel if provided to metadata
+#Apply relevel if provided; this must be done before calling DESeq()
 if (isProvided(params$path_relevel)) {
     relevel_table <- read.table(params$path_relevel, sep="\t", header = T, colClasses = "character")
     write.table(relevel_table, file="differential_gene_expression/metadata/relevel.tsv")
@@ -553,6 +550,9 @@ if (isProvided(params$path_relevel)) {
     }
 }
 
+# Run actual DESeq analysis
+cds <- DESeq(cds)
+
 # sizeFactors(cds) as indicator of library sequencing depth
 if (params$input_type == "featurecounts") {
     sf <- sizeFactors(cds)

From 691bcc47f5de9d19713776fe7508a9356fca7543 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 3 Jul 2023 08:53:46 +0200
Subject: [PATCH 069/124] nf-core lint

---
 lib/NfcoreTemplate.groovy | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
index 2023f451..000bb929 100755
--- a/lib/NfcoreTemplate.groovy
+++ b/lib/NfcoreTemplate.groovy
@@ -128,7 +128,7 @@ class NfcoreTemplate {
         def email_html    = html_template.toString()
 
         // Render the sendmail template
-        def max_multiqc_email_size = params.max_multiqc_email_size as nextflow.util.MemoryUnit
+        def max_multiqc_email_size = (params.containsKey('max_multiqc_email_size') ? params.max_multiqc_email_size : 0) as nextflow.util.MemoryUnit
         def smail_fields           = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ]
         def sf                     = new File("$projectDir/assets/sendmail_template.txt")
         def sendmail_template      = engine.createTemplate(sf).make(smail_fields)

From 1229e8b8132d7132ad127742f4eb2e919a9046d1 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 3 Jul 2023 15:09:28 +0200
Subject: [PATCH 070/124] updated test.yml

---
 tests/test_relevel.yml | 30 +++++++++++++++---------------
 1 file changed, 15 insertions(+), 15 deletions(-)

diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml
index 92e927d6..885da98c 100644
--- a/tests/test_relevel.yml
+++ b/tests/test_relevel.yml
@@ -3,12 +3,12 @@
   tags:
     - test_relevel
   files:
-    - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_genotype_WT_vs_KO.tsv
-      md5sum: 93fd663cb976c102144e1ffb34d88e53
+    - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_genotype_KO_vs_WT.tsv
+      md5sum: d0590d6438ce02b88c58c8c9721b0740
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_Treated_vs_Control.tsv
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
-      md5sum: 81d3437334849b3b4de4de6729a3d54d
+      md5sum: 6b8731f514a7e7c1645376b0ffde1a6d
     - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
     - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
@@ -22,18 +22,18 @@
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL11_ENSMUSG00000004371.pdf
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL11_ENSMUSG00000004371.png
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL11_ENSMUSG00000004371.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_KO_vs_WT.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_KO_vs_WT.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_genotype_KO_vs_WT.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_Treated_vs_Control.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_Treated_vs_Control.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_KO_vs_WT.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_KO_vs_WT.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_genotype_KO_vs_WT.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_Treated_vs_Control.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_Treated_vs_Control.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_Treated_vs_Control.svg
@@ -43,15 +43,15 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_Treated_vs_Control.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_Treated_vs_Control.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_Treated_vs_Control.svg

From f59f142936bcdf032dc5d77d59d0a169ec39a0d3 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 3 Jul 2023 15:32:35 +0200
Subject: [PATCH 071/124] changed md5sum

---
 tests/test_relevel.yml | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml
index 885da98c..ccec3ac7 100644
--- a/tests/test_relevel.yml
+++ b/tests/test_relevel.yml
@@ -4,7 +4,7 @@
     - test_relevel
   files:
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_genotype_KO_vs_WT.tsv
-      md5sum: d0590d6438ce02b88c58c8c9721b0740
+      md5sum: 02380f76fff2658c76cff0337fad07c9
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_Treated_vs_Control.tsv
       md5sum: cb1f49e7fe93b4ef2673eb99fd8793cd
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv

From 1bc3887e84f3789d4d219d1ba9a2d333b32dbedf Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 11 Jul 2023 14:37:13 +0200
Subject: [PATCH 072/124] Changed filenames of histograms of p values

---
 assets/RNAseq_report.Rmd       |  6 +++---
 tests/test.yml                 | 12 ++++++------
 tests/test_batcheffect.yml     | 12 ++++++------
 tests/test_contrast_list.yml   | 12 ++++++------
 tests/test_contrast_matrix.yml | 12 ++++++------
 tests/test_custom_gmt.yml      | 12 ++++++------
 tests/test_full.yml            | 12 ++++++------
 tests/test_no_multiqc.yml      | 12 ++++++------
 tests/test_relevel.yml         | 12 ++++++------
 tests/test_star_rsem.yml       | 18 +++++++++---------
 tests/test_star_salmon.yml     | 18 +++++++++---------
 tests/test_use_vst.yml         | 12 ++++++------
 12 files changed, 75 insertions(+), 75 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 1c73c05f..67c6201f 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -748,21 +748,21 @@ for (i in resultsNames(cds)[-1]) {
     h1 <- hist(res$pvalue[!use], breaks=0:50/50, plot=FALSE)
     h2 <- hist(res$pvalue[use], breaks=0:50/50, plot=FALSE)
     colori <- c('do not pass'="khaki", 'pass'="powderblue")
-    pdf(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".pdf",sep=""))
+    pdf(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_",i,".pdf",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
             col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
         adj = c(0.5,1.7), xpd=NA)
     legend("topleft", fill=rev(colori), legend=rev(names(colori)))
     dev.off()
-    png(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".png",sep=""))
+    png(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_",i,".png",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
             col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
         adj = c(0.5,1.7), xpd=NA)
     legend("topleft", fill=rev(colori), legend=rev(names(colori)))
     dev.off()
-    svg(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values",i,".svg",sep=""))
+    svg(paste("differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_",i,".svg",sep=""))
     barplot(height = rbind(h1$density, h2$density), beside = FALSE,
             col = colori, space = 0, main = "", xlab="p-value",ylab="frequency")
     text(x = c(0, length(h1$counts)), y = 0, label = paste(c(0,1)),
diff --git a/tests/test.yml b/tests/test.yml
index 2adebac0..56ff217d 100644
--- a/tests/test.yml
+++ b/tests/test.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml
index 72150e5b..08511fa9 100644
--- a/tests/test_batcheffect.yml
+++ b/tests/test_batcheffect.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_contrast_list.yml b/tests/test_contrast_list.yml
index f3aebcd6..e090ef90 100644
--- a/tests/test_contrast_list.yml
+++ b/tests/test_contrast_list.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_contrast_matrix.yml b/tests/test_contrast_matrix.yml
index 05d5a02b..a9059ea0 100644
--- a/tests/test_contrast_matrix.yml
+++ b/tests/test_contrast_matrix.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml
index 071c297b..589ccf13 100644
--- a/tests/test_custom_gmt.yml
+++ b/tests/test_custom_gmt.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_full.yml b/tests/test_full.yml
index 21be0ef4..c4477cbf 100644
--- a/tests/test_full.yml
+++ b/tests/test_full.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_no_multiqc.yml b/tests/test_no_multiqc.yml
index 9e86ef4d..417a0f5c 100644
--- a/tests/test_no_multiqc.yml
+++ b/tests/test_no_multiqc.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg
diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml
index ccec3ac7..8af64813 100644
--- a/tests/test_relevel.yml
+++ b/tests/test_relevel.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_KO_vs_WT.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_KO_vs_WT.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_KO_vs_WT.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_KO_vs_WT.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_KO_vs_WT.svg
diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml
index e55c497e..c9cbdd49 100644
--- a/tests/test_star_rsem.yml
+++ b/tests/test_star_rsem.yml
@@ -52,15 +52,15 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.svg
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index 3a8dc898..20b498cc 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -28,15 +28,15 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_H1_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_K562_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_cellline_MCF7_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_K562_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_MCF7_vs_GM12878.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_cellline_H1_vs_GM12878.svg
diff --git a/tests/test_use_vst.yml b/tests/test_use_vst.yml
index eb8a731a..ec6efa1e 100644
--- a/tests/test_use_vst.yml
+++ b/tests/test_use_vst.yml
@@ -43,12 +43,12 @@
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_genotype_WT_vs_KO.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.valuescondition_treatment_Treated_vs_Control.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_genotype_WT_vs_KO.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_Treated_vs_Control.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_genotype_WT_vs_KO.svg

From d77e63c27f9ad746a3ce2890f175b3bdca92fe99 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 14 Jul 2023 13:05:41 +0200
Subject: [PATCH 073/124] Initial template commit

---
 .devcontainer/devcontainer.json               |  27 ++
 .editorconfig                                 |  24 +
 .gitattributes                                |   4 +
 .github/.dockstore.yml                        |   6 +
 .github/CONTRIBUTING.md                       | 118 +++++
 .github/ISSUE_TEMPLATE/bug_report.yml         |  50 ++
 .github/ISSUE_TEMPLATE/config.yml             |   7 +
 .github/ISSUE_TEMPLATE/feature_request.yml    |  11 +
 .github/PULL_REQUEST_TEMPLATE.md              |  25 +
 .github/workflows/awsfulltest.yml             |  39 ++
 .github/workflows/awstest.yml                 |  33 ++
 .github/workflows/branch.yml                  |  44 ++
 .github/workflows/ci.yml                      |  43 ++
 .github/workflows/clean-up.yml                |  24 +
 .github/workflows/fix-linting.yml             |  55 +++
 .github/workflows/linting.yml                 | 108 +++++
 .github/workflows/linting_comment.yml         |  28 ++
 .gitignore                                    |   8 +
 .gitpod.yml                                   |  19 +
 .nf-core.yml                                  |   1 +
 .pre-commit-config.yaml                       |   5 +
 .prettierignore                               |  12 +
 .prettierrc.yml                               |   1 +
 CHANGELOG.md                                  |  16 +
 CITATIONS.md                                  |  41 ++
 CODE_OF_CONDUCT.md                            | 111 +++++
 LICENSE                                       |  21 +
 README.md                                     | 106 +++++
 assets/adaptivecard.json                      |  67 +++
 assets/email_template.html                    |  53 +++
 assets/email_template.txt                     |  39 ++
 assets/methods_description_template.yml       |  29 ++
 assets/multiqc_config.yml                     |  13 +
 assets/nf-core-rnadeseq_logo_light.png        | Bin 0 -> 75779 bytes
 assets/samplesheet.csv                        |   3 +
 assets/schema_input.json                      |  36 ++
 assets/sendmail_template.txt                  |  53 +++
 assets/slackreport.json                       |  34 ++
 bin/check_samplesheet.py                      | 259 +++++++++++
 conf/base.config                              |  65 +++
 conf/igenomes.config                          | 440 ++++++++++++++++++
 conf/modules.config                           |  41 ++
 conf/test.config                              |  29 ++
 conf/test_full.config                         |  24 +
 docs/README.md                                |  10 +
 docs/images/mqc_fastqc_adapter.png            | Bin 0 -> 23458 bytes
 docs/images/mqc_fastqc_counts.png             | Bin 0 -> 33918 bytes
 docs/images/mqc_fastqc_quality.png            | Bin 0 -> 55769 bytes
 docs/images/nf-core-rnadeseq_logo_dark.png    | Bin 0 -> 75896 bytes
 docs/images/nf-core-rnadeseq_logo_light.png   | Bin 0 -> 75779 bytes
 docs/output.md                                |  68 +++
 docs/usage.md                                 | 216 +++++++++
 lib/NfcoreTemplate.groovy                     | 336 +++++++++++++
 lib/Utils.groovy                              |  47 ++
 lib/WorkflowMain.groovy                       |  63 +++
 lib/WorkflowRnadeseq.groovy                   | 122 +++++
 lib/nfcore_external_java_deps.jar             | Bin 0 -> 2291171 bytes
 main.nf                                       |  79 ++++
 modules.json                                  |  27 ++
 modules/local/samplesheet_check.nf            |  31 ++
 .../custom/dumpsoftwareversions/main.nf       |  24 +
 .../custom/dumpsoftwareversions/meta.yml      |  36 ++
 .../templates/dumpsoftwareversions.py         | 102 ++++
 modules/nf-core/fastqc/main.nf                |  51 ++
 modules/nf-core/fastqc/meta.yml               |  52 +++
 modules/nf-core/multiqc/main.nf               |  53 +++
 modules/nf-core/multiqc/meta.yml              |  56 +++
 nextflow.config                               | 269 +++++++++++
 nextflow_schema.json                          | 303 ++++++++++++
 pyproject.toml                                |  10 +
 subworkflows/local/input_check.nf             |  44 ++
 tower.yml                                     |   5 +
 workflows/rnadeseq.nf                         | 133 ++++++
 73 files changed, 4309 insertions(+)
 create mode 100644 .devcontainer/devcontainer.json
 create mode 100644 .editorconfig
 create mode 100644 .gitattributes
 create mode 100644 .github/.dockstore.yml
 create mode 100644 .github/CONTRIBUTING.md
 create mode 100644 .github/ISSUE_TEMPLATE/bug_report.yml
 create mode 100644 .github/ISSUE_TEMPLATE/config.yml
 create mode 100644 .github/ISSUE_TEMPLATE/feature_request.yml
 create mode 100644 .github/PULL_REQUEST_TEMPLATE.md
 create mode 100644 .github/workflows/awsfulltest.yml
 create mode 100644 .github/workflows/awstest.yml
 create mode 100644 .github/workflows/branch.yml
 create mode 100644 .github/workflows/ci.yml
 create mode 100644 .github/workflows/clean-up.yml
 create mode 100644 .github/workflows/fix-linting.yml
 create mode 100644 .github/workflows/linting.yml
 create mode 100644 .github/workflows/linting_comment.yml
 create mode 100644 .gitignore
 create mode 100644 .gitpod.yml
 create mode 100644 .nf-core.yml
 create mode 100644 .pre-commit-config.yaml
 create mode 100644 .prettierignore
 create mode 100644 .prettierrc.yml
 create mode 100644 CHANGELOG.md
 create mode 100644 CITATIONS.md
 create mode 100644 CODE_OF_CONDUCT.md
 create mode 100644 LICENSE
 create mode 100644 README.md
 create mode 100644 assets/adaptivecard.json
 create mode 100644 assets/email_template.html
 create mode 100644 assets/email_template.txt
 create mode 100644 assets/methods_description_template.yml
 create mode 100644 assets/multiqc_config.yml
 create mode 100644 assets/nf-core-rnadeseq_logo_light.png
 create mode 100644 assets/samplesheet.csv
 create mode 100644 assets/schema_input.json
 create mode 100644 assets/sendmail_template.txt
 create mode 100644 assets/slackreport.json
 create mode 100755 bin/check_samplesheet.py
 create mode 100644 conf/base.config
 create mode 100644 conf/igenomes.config
 create mode 100644 conf/modules.config
 create mode 100644 conf/test.config
 create mode 100644 conf/test_full.config
 create mode 100644 docs/README.md
 create mode 100755 docs/images/mqc_fastqc_adapter.png
 create mode 100755 docs/images/mqc_fastqc_counts.png
 create mode 100755 docs/images/mqc_fastqc_quality.png
 create mode 100644 docs/images/nf-core-rnadeseq_logo_dark.png
 create mode 100644 docs/images/nf-core-rnadeseq_logo_light.png
 create mode 100644 docs/output.md
 create mode 100644 docs/usage.md
 create mode 100755 lib/NfcoreTemplate.groovy
 create mode 100644 lib/Utils.groovy
 create mode 100755 lib/WorkflowMain.groovy
 create mode 100755 lib/WorkflowRnadeseq.groovy
 create mode 100644 lib/nfcore_external_java_deps.jar
 create mode 100644 main.nf
 create mode 100644 modules.json
 create mode 100644 modules/local/samplesheet_check.nf
 create mode 100644 modules/nf-core/custom/dumpsoftwareversions/main.nf
 create mode 100644 modules/nf-core/custom/dumpsoftwareversions/meta.yml
 create mode 100755 modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
 create mode 100644 modules/nf-core/fastqc/main.nf
 create mode 100644 modules/nf-core/fastqc/meta.yml
 create mode 100644 modules/nf-core/multiqc/main.nf
 create mode 100644 modules/nf-core/multiqc/meta.yml
 create mode 100644 nextflow.config
 create mode 100644 nextflow_schema.json
 create mode 100644 pyproject.toml
 create mode 100644 subworkflows/local/input_check.nf
 create mode 100644 tower.yml
 create mode 100644 workflows/rnadeseq.nf

diff --git a/.devcontainer/devcontainer.json b/.devcontainer/devcontainer.json
new file mode 100644
index 00000000..ea27a584
--- /dev/null
+++ b/.devcontainer/devcontainer.json
@@ -0,0 +1,27 @@
+{
+    "name": "nfcore",
+    "image": "nfcore/gitpod:latest",
+    "remoteUser": "gitpod",
+
+    // Configure tool-specific properties.
+    "customizations": {
+        // Configure properties specific to VS Code.
+        "vscode": {
+            // Set *default* container specific settings.json values on container create.
+            "settings": {
+                "python.defaultInterpreterPath": "/opt/conda/bin/python",
+                "python.linting.enabled": true,
+                "python.linting.pylintEnabled": true,
+                "python.formatting.autopep8Path": "/opt/conda/bin/autopep8",
+                "python.formatting.yapfPath": "/opt/conda/bin/yapf",
+                "python.linting.flake8Path": "/opt/conda/bin/flake8",
+                "python.linting.pycodestylePath": "/opt/conda/bin/pycodestyle",
+                "python.linting.pydocstylePath": "/opt/conda/bin/pydocstyle",
+                "python.linting.pylintPath": "/opt/conda/bin/pylint"
+            },
+
+            // Add the IDs of extensions you want installed when the container is created.
+            "extensions": ["ms-python.python", "ms-python.vscode-pylance", "nf-core.nf-core-extensionpack"]
+        }
+    }
+}
diff --git a/.editorconfig b/.editorconfig
new file mode 100644
index 00000000..b6b31907
--- /dev/null
+++ b/.editorconfig
@@ -0,0 +1,24 @@
+root = true
+
+[*]
+charset = utf-8
+end_of_line = lf
+insert_final_newline = true
+trim_trailing_whitespace = true
+indent_size = 4
+indent_style = space
+
+[*.{md,yml,yaml,html,css,scss,js}]
+indent_size = 2
+
+# These files are edited and tested upstream in nf-core/modules
+[/modules/nf-core/**]
+charset = unset
+end_of_line = unset
+insert_final_newline = unset
+trim_trailing_whitespace = unset
+indent_style = unset
+indent_size = unset
+
+[/assets/email*]
+indent_size = unset
diff --git a/.gitattributes b/.gitattributes
new file mode 100644
index 00000000..7a2dabc2
--- /dev/null
+++ b/.gitattributes
@@ -0,0 +1,4 @@
+*.config linguist-language=nextflow
+*.nf.test linguist-language=nextflow
+modules/nf-core/** linguist-generated
+subworkflows/nf-core/** linguist-generated
diff --git a/.github/.dockstore.yml b/.github/.dockstore.yml
new file mode 100644
index 00000000..191fabd2
--- /dev/null
+++ b/.github/.dockstore.yml
@@ -0,0 +1,6 @@
+# Dockstore config version, not pipeline version
+version: 1.2
+workflows:
+  - subclass: nfl
+    primaryDescriptorPath: /nextflow.config
+    publish: True
diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md
new file mode 100644
index 00000000..ebec4799
--- /dev/null
+++ b/.github/CONTRIBUTING.md
@@ -0,0 +1,118 @@
+# nf-core/rnadeseq: Contributing Guidelines
+
+Hi there!
+Many thanks for taking an interest in improving nf-core/rnadeseq.
+
+We try to manage the required tasks for nf-core/rnadeseq using GitHub issues, you probably came to this page when creating one.
+Please use the pre-filled template to save time.
+
+However, don't be put off by this template - other more general issues and suggestions are welcome!
+Contributions to the code are even more welcome ;)
+
+> If you need help using or modifying nf-core/rnadeseq then the best place to ask is on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)).
+
+## Contribution workflow
+
+If you'd like to write some code for nf-core/rnadeseq, the standard workflow is as follows:
+
+1. Check that there isn't already an issue about your idea in the [nf-core/rnadeseq issues](https://github.com/nf-core/rnadeseq/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this
+2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/rnadeseq repository](https://github.com/nf-core/rnadeseq) to your GitHub account
+3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions)
+4. Use `nf-core schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10).
+5. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged
+
+If you're not used to this workflow with git, you can start with some [docs from GitHub](https://help.github.com/en/github/collaborating-with-issues-and-pull-requests) or even their [excellent `git` resources](https://try.github.io/).
+
+## Tests
+
+When you create a pull request with changes, [GitHub Actions](https://github.com/features/actions) will run automatic tests.
+Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then.
+
+There are typically two types of tests that run:
+
+### Lint tests
+
+`nf-core` has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to.
+To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint <pipeline-directory>` command.
+
+If any failures or warnings are encountered, please follow the listed URL for more documentation.
+
+### Pipeline tests
+
+Each `nf-core` pipeline should be set up with a minimal set of test-data.
+`GitHub Actions` then runs the pipeline on this data to ensure that it exits successfully.
+If there are any failures then the automated tests fail.
+These tests are run both with the latest available version of `Nextflow` and also the minimum required version that is stated in the pipeline code.
+
+## Patch
+
+:warning: Only in the unlikely and regretful event of a release happening with a bug.
+
+- On your own fork, make a new branch `patch` based on `upstream/master`.
+- Fix the bug, and bump version (X.Y.Z+1).
+- A PR should be made on `master` from patch to directly this particular bug.
+
+## Getting help
+
+For further information/help, please consult the [nf-core/rnadeseq documentation](https://nf-co.re/rnadeseq/usage) and don't hesitate to get in touch on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)).
+
+## Pipeline contribution conventions
+
+To make the nf-core/rnadeseq code and processing logic more understandable for new contributors and to ensure quality, we semi-standardise the way the code and other contributions are written.
+
+### Adding a new step
+
+If you wish to contribute a new step, please use the following coding standards:
+
+1. Define the corresponding input channel into your new process from the expected previous process channel
+2. Write the process block (see below).
+3. Define the output channel if needed (see below).
+4. Add any new parameters to `nextflow.config` with a default (see below).
+5. Add any new parameters to `nextflow_schema.json` with help text (via the `nf-core schema build` tool).
+6. Add sanity checks and validation for all relevant parameters.
+7. Perform local tests to validate that the new code works as expected.
+8. If applicable, add a new test command in `.github/workflow/ci.yml`.
+9. Update MultiQC config `assets/multiqc_config.yml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module.
+10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`.
+
+### Default values
+
+Parameters should be initialised / defined with default values in `nextflow.config` under the `params` scope.
+
+Once there, use `nf-core schema build` to add to `nextflow_schema.json`.
+
+### Default processes resource requirements
+
+Sensible defaults for process resource requirements (CPUs / memory / time) for a process should be defined in `conf/base.config`. These should generally be specified generic with `withLabel:` selectors so they can be shared across multiple processes/steps of the pipeline. A nf-core standard set of labels that should be followed where possible can be seen in the [nf-core pipeline template](https://github.com/nf-core/tools/blob/master/nf_core/pipeline-template/conf/base.config), which has the default process as a single core-process, and then different levels of multi-core configurations for increasingly large memory requirements defined with standardised labels.
+
+The process resources can be passed on to the tool dynamically within the process with the `${task.cpu}` and `${task.memory}` variables in the `script:` block.
+
+### Naming schemes
+
+Please use the following naming schemes, to make it easy to understand what is going where.
+
+- initial process channel: `ch_output_from_<process>`
+- intermediate and terminal channels: `ch_<previousprocess>_for_<nextprocess>`
+
+### Nextflow version bumping
+
+If you are using a new feature from core Nextflow, you may bump the minimum required version of nextflow in the pipeline with: `nf-core bump-version --nextflow . [min-nf-version]`
+
+### Images and figures
+
+For overview images and other documents we follow the nf-core [style guidelines and examples](https://nf-co.re/developers/design_guidelines).
+
+## GitHub Codespaces
+
+This repo includes a devcontainer configuration which will create a GitHub Codespaces for Nextflow development! This is an online developer environment that runs in your browser, complete with VSCode and a terminal.
+
+To get started:
+
+- Open the repo in [Codespaces](https://github.com/nf-core/rnadeseq/codespaces)
+- Tools installed
+  - nf-core
+  - Nextflow
+
+Devcontainer specs:
+
+- [DevContainer config](.devcontainer/devcontainer.json)
diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml
new file mode 100644
index 00000000..6e8602bd
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/bug_report.yml
@@ -0,0 +1,50 @@
+name: Bug report
+description: Report something that is broken or incorrect
+labels: bug
+body:
+  - type: markdown
+    attributes:
+      value: |
+        Before you post this issue, please check the documentation:
+
+        - [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting)
+        - [nf-core/rnadeseq pipeline documentation](https://nf-co.re/rnadeseq/usage)
+
+  - type: textarea
+    id: description
+    attributes:
+      label: Description of the bug
+      description: A clear and concise description of what the bug is.
+    validations:
+      required: true
+
+  - type: textarea
+    id: command_used
+    attributes:
+      label: Command used and terminal output
+      description: Steps to reproduce the behaviour. Please paste the command you used to launch the pipeline and the output from your terminal.
+      render: console
+      placeholder: |
+        $ nextflow run ...
+
+        Some output where something broke
+
+  - type: textarea
+    id: files
+    attributes:
+      label: Relevant files
+      description: |
+        Please drag and drop the relevant files here. Create a `.zip` archive if the extension is not allowed.
+        Your verbose log file `.nextflow.log` is often useful _(this is a hidden file in the directory where you launched the pipeline)_ as well as custom Nextflow configuration files.
+
+  - type: textarea
+    id: system
+    attributes:
+      label: System information
+      description: |
+        * Nextflow version _(eg. 23.04.0)_
+        * Hardware _(eg. HPC, Desktop, Cloud)_
+        * Executor _(eg. slurm, local, awsbatch)_
+        * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, or Apptainer)_
+        * OS _(eg. CentOS Linux, macOS, Linux Mint)_
+        * Version of nf-core/rnadeseq _(eg. 1.1, 1.5, 1.8.2)_
diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml
new file mode 100644
index 00000000..1c032cae
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/config.yml
@@ -0,0 +1,7 @@
+contact_links:
+  - name: Join nf-core
+    url: https://nf-co.re/join
+    about: Please join the nf-core community here
+  - name: "Slack #rnadeseq channel"
+    url: https://nfcore.slack.com/channels/rnadeseq
+    about: Discussion about the nf-core/rnadeseq pipeline
diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml
new file mode 100644
index 00000000..6bd51f6c
--- /dev/null
+++ b/.github/ISSUE_TEMPLATE/feature_request.yml
@@ -0,0 +1,11 @@
+name: Feature request
+description: Suggest an idea for the nf-core/rnadeseq pipeline
+labels: enhancement
+body:
+  - type: textarea
+    id: description
+    attributes:
+      label: Description of feature
+      description: Please describe your suggestion for a new feature. It might help to describe a problem or use case, plus any alternatives that you have considered.
+    validations:
+      required: true
diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md
new file mode 100644
index 00000000..6d057bf9
--- /dev/null
+++ b/.github/PULL_REQUEST_TEMPLATE.md
@@ -0,0 +1,25 @@
+<!--
+# nf-core/rnadeseq pull request
+
+Many thanks for contributing to nf-core/rnadeseq!
+
+Please fill in the appropriate checklist below (delete whatever is not relevant).
+These are the most common things requested on pull requests (PRs).
+
+Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release.
+
+Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/rnadeseq/tree/master/.github/CONTRIBUTING.md)
+-->
+
+## PR checklist
+
+- [ ] This comment contains a description of changes (with reason).
+- [ ] If you've fixed a bug or added code that should be tested, add tests!
+- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/rnadeseq/tree/master/.github/CONTRIBUTING.md)
+- [ ] If necessary, also make a PR on the nf-core/rnadeseq _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository.
+- [ ] Make sure your code lints (`nf-core lint`).
+- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir <OUTDIR>`).
+- [ ] Usage Documentation in `docs/usage.md` is updated.
+- [ ] Output Documentation in `docs/output.md` is updated.
+- [ ] `CHANGELOG.md` is updated.
+- [ ] `README.md` is updated (including new tool citations and authors/contributors).
diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml
new file mode 100644
index 00000000..84743a89
--- /dev/null
+++ b/.github/workflows/awsfulltest.yml
@@ -0,0 +1,39 @@
+name: nf-core AWS full size tests
+# This workflow is triggered on published releases.
+# It can be additionally triggered manually with GitHub actions workflow dispatch button.
+# It runs the -profile 'test_full' on AWS batch
+
+on:
+  release:
+    types: [published]
+  workflow_dispatch:
+jobs:
+  run-tower:
+    name: Run AWS full tests
+    if: github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      - name: Launch workflow via tower
+        uses: seqeralabs/action-tower-launch@v2
+        # TODO nf-core: You can customise AWS full pipeline tests as required
+        # Add full size test data (but still relatively small datasets for few samples)
+        # on the `test_full.config` test runs with only one set of parameters
+        with:
+          workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }}
+          access_token: ${{ secrets.TOWER_ACCESS_TOKEN }}
+          compute_env: ${{ secrets.TOWER_COMPUTE_ENV }}
+          revision: ${{ github.sha }}
+          workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }}
+          parameters: |
+            {
+              "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}",
+              "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-${{ github.sha }}"
+            }
+          profiles: test_full
+
+      - uses: actions/upload-artifact@v3
+        with:
+          name: Tower debug log file
+          path: |
+            tower_action_*.log
+            tower_action_*.json
diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml
new file mode 100644
index 00000000..a1300168
--- /dev/null
+++ b/.github/workflows/awstest.yml
@@ -0,0 +1,33 @@
+name: nf-core AWS test
+# This workflow can be triggered manually with the GitHub actions workflow dispatch button.
+# It runs the -profile 'test' on AWS batch
+
+on:
+  workflow_dispatch:
+jobs:
+  run-tower:
+    name: Run AWS tests
+    if: github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      # Launch workflow using Tower CLI tool action
+      - name: Launch workflow via tower
+        uses: seqeralabs/action-tower-launch@v2
+        with:
+          workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }}
+          access_token: ${{ secrets.TOWER_ACCESS_TOKEN }}
+          compute_env: ${{ secrets.TOWER_COMPUTE_ENV }}
+          revision: ${{ github.sha }}
+          workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }}
+          parameters: |
+            {
+              "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-test-${{ github.sha }}"
+            }
+          profiles: test
+
+      - uses: actions/upload-artifact@v3
+        with:
+          name: Tower debug log file
+          path: |
+            tower_action_*.log
+            tower_action_*.json
diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml
new file mode 100644
index 00000000..8230e02e
--- /dev/null
+++ b/.github/workflows/branch.yml
@@ -0,0 +1,44 @@
+name: nf-core branch protection
+# This workflow is triggered on PRs to master branch on the repository
+# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev`
+on:
+  pull_request_target:
+    branches: [master]
+
+jobs:
+  test:
+    runs-on: ubuntu-latest
+    steps:
+      # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches
+      - name: Check PRs
+        if: github.repository == 'nf-core/rnadeseq'
+        run: |
+          { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]
+
+      # If the above check failed, post a comment on the PR explaining the failure
+      # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets
+      - name: Post PR comment
+        if: failure()
+        uses: mshick/add-pr-comment@v1
+        with:
+          message: |
+            ## This PR is against the `master` branch :x:
+
+            * Do not close this PR
+            * Click _Edit_ and change the `base` to `dev`
+            * This CI test will remain failed until you push a new commit
+
+            ---
+
+            Hi @${{ github.event.pull_request.user.login }},
+
+            It looks like this pull-request is has been made against the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `master` branch.
+            The `master` branch on nf-core repositories should always contain code from the latest release.
+            Because of this, PRs to `master` are only allowed if they come from the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `dev` branch.
+
+            You do not need to close this PR, you can change the target branch to `dev` by clicking the _"Edit"_ button at the top of this page.
+            Note that even after this, the test will continue to show as failing until you push a new commit.
+
+            Thanks again for your contribution!
+          repo-token: ${{ secrets.GITHUB_TOKEN }}
+          allow-repeats: false
diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
new file mode 100644
index 00000000..70772968
--- /dev/null
+++ b/.github/workflows/ci.yml
@@ -0,0 +1,43 @@
+name: nf-core CI
+# This workflow runs the pipeline with the minimal test dataset to check that it completes without any syntax errors
+on:
+  push:
+    branches:
+      - dev
+  pull_request:
+  release:
+    types: [published]
+
+env:
+  NXF_ANSI_LOG: false
+
+concurrency:
+  group: "${{ github.workflow }}-${{ github.event.pull_request.number || github.ref }}"
+  cancel-in-progress: true
+
+jobs:
+  test:
+    name: Run pipeline with test data
+    # Only run on push if this is the nf-core dev branch (merged PRs)
+    if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/rnadeseq') }}"
+    runs-on: ubuntu-latest
+    strategy:
+      matrix:
+        NXF_VER:
+          - "23.04.0"
+          - "latest-everything"
+    steps:
+      - name: Check out pipeline code
+        uses: actions/checkout@v3
+
+      - name: Install Nextflow
+        uses: nf-core/setup-nextflow@v1
+        with:
+          version: "${{ matrix.NXF_VER }}"
+
+      - name: Run pipeline with test data
+        # TODO nf-core: You can customise CI pipeline run tests as required
+        # For example: adding multiple test runs with different parameters
+        # Remember that you can parallelise this by using strategy.matrix
+        run: |
+          nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results
diff --git a/.github/workflows/clean-up.yml b/.github/workflows/clean-up.yml
new file mode 100644
index 00000000..694e90ec
--- /dev/null
+++ b/.github/workflows/clean-up.yml
@@ -0,0 +1,24 @@
+name: "Close user-tagged issues and PRs"
+on:
+  schedule:
+    - cron: "0 0 * * 0" # Once a week
+
+jobs:
+  clean-up:
+    runs-on: ubuntu-latest
+    permissions:
+      issues: write
+      pull-requests: write
+    steps:
+      - uses: actions/stale@v7
+        with:
+          stale-issue-message: "This issue has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment otherwise this issue will be closed in 20 days."
+          stale-pr-message: "This PR has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment if it is still useful."
+          close-issue-message: "This issue was closed because it has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor and then staled for 20 days with no activity."
+          days-before-stale: 30
+          days-before-close: 20
+          days-before-pr-close: -1
+          any-of-labels: "awaiting-changes,awaiting-feedback"
+          exempt-issue-labels: "WIP"
+          exempt-pr-labels: "WIP"
+          repo-token: "${{ secrets.GITHUB_TOKEN }}"
diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml
new file mode 100644
index 00000000..588b4f43
--- /dev/null
+++ b/.github/workflows/fix-linting.yml
@@ -0,0 +1,55 @@
+name: Fix linting from a comment
+on:
+  issue_comment:
+    types: [created]
+
+jobs:
+  deploy:
+    # Only run if comment is on a PR with the main repo, and if it contains the magic keywords
+    if: >
+      contains(github.event.comment.html_url, '/pull/') &&
+      contains(github.event.comment.body, '@nf-core-bot fix linting') &&
+      github.repository == 'nf-core/rnadeseq'
+    runs-on: ubuntu-latest
+    steps:
+      # Use the @nf-core-bot token to check out so we can push later
+      - uses: actions/checkout@v3
+        with:
+          token: ${{ secrets.nf_core_bot_auth_token }}
+
+      # Action runs on the issue comment, so we don't get the PR by default
+      # Use the gh cli to check out the PR
+      - name: Checkout Pull Request
+        run: gh pr checkout ${{ github.event.issue.number }}
+        env:
+          GITHUB_TOKEN: ${{ secrets.nf_core_bot_auth_token }}
+
+      - uses: actions/setup-node@v3
+
+      - name: Install Prettier
+        run: npm install -g prettier @prettier/plugin-php
+
+      # Check that we actually need to fix something
+      - name: Run 'prettier --check'
+        id: prettier_status
+        run: |
+          if prettier --check ${GITHUB_WORKSPACE}; then
+            echo "result=pass" >> $GITHUB_OUTPUT
+          else
+            echo "result=fail" >> $GITHUB_OUTPUT
+          fi
+
+      - name: Run 'prettier --write'
+        if: steps.prettier_status.outputs.result == 'fail'
+        run: prettier --write ${GITHUB_WORKSPACE}
+
+      - name: Commit & push changes
+        if: steps.prettier_status.outputs.result == 'fail'
+        run: |
+          git config user.email "core@nf-co.re"
+          git config user.name "nf-core-bot"
+          git config push.default upstream
+          git add .
+          git status
+          git commit -m "[automated] Fix linting with Prettier"
+          git push
diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml
new file mode 100644
index 00000000..888cb4bc
--- /dev/null
+++ b/.github/workflows/linting.yml
@@ -0,0 +1,108 @@
+name: nf-core linting
+# This workflow is triggered on pushes and PRs to the repository.
+# It runs the `nf-core lint` and markdown lint tests to ensure
+# that the code meets the nf-core guidelines.
+on:
+  push:
+    branches:
+      - dev
+  pull_request:
+  release:
+    types: [published]
+
+jobs:
+  EditorConfig:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - uses: actions/setup-node@v3
+
+      - name: Install editorconfig-checker
+        run: npm install -g editorconfig-checker
+
+      - name: Run ECLint check
+        run: editorconfig-checker -exclude README.md $(find .* -type f | grep -v '.git\|.py\|.md\|json\|yml\|yaml\|html\|css\|work\|.nextflow\|build\|nf_core.egg-info\|log.txt\|Makefile')
+
+  Prettier:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - uses: actions/setup-node@v3
+
+      - name: Install Prettier
+        run: npm install -g prettier
+
+      - name: Run Prettier --check
+        run: prettier --check ${GITHUB_WORKSPACE}
+
+  PythonBlack:
+    runs-on: ubuntu-latest
+    steps:
+      - uses: actions/checkout@v3
+
+      - name: Check code lints with Black
+        uses: psf/black@stable
+
+      # If the above check failed, post a comment on the PR explaining the failure
+      - name: Post PR comment
+        if: failure()
+        uses: mshick/add-pr-comment@v1
+        with:
+          message: |
+            ## Python linting (`black`) is failing
+
+            To keep the code consistent with lots of contributors, we run automated code consistency checks.
+            To fix this CI test, please run:
+
+            * Install [`black`](https://black.readthedocs.io/en/stable/): `pip install black`
+            * Fix formatting errors in your pipeline: `black .`
+
+            Once you push these changes the test should pass, and you can hide this comment :+1:
+
+            We highly recommend setting up Black in your code editor so that this formatting is done automatically on save. Ask about it on Slack for help!
+
+            Thanks again for your contribution!
+          repo-token: ${{ secrets.GITHUB_TOKEN }}
+          allow-repeats: false
+
+  nf-core:
+    runs-on: ubuntu-latest
+    steps:
+      - name: Check out pipeline code
+        uses: actions/checkout@v3
+
+      - name: Install Nextflow
+        uses: nf-core/setup-nextflow@v1
+
+      - uses: actions/setup-python@v4
+        with:
+          python-version: "3.8"
+          architecture: "x64"
+
+      - name: Install dependencies
+        run: |
+          python -m pip install --upgrade pip
+          pip install nf-core
+
+      - name: Run nf-core lint
+        env:
+          GITHUB_COMMENTS_URL: ${{ github.event.pull_request.comments_url }}
+          GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }}
+          GITHUB_PR_COMMIT: ${{ github.event.pull_request.head.sha }}
+        run: nf-core -l lint_log.txt lint --dir ${GITHUB_WORKSPACE} --markdown lint_results.md
+
+      - name: Save PR number
+        if: ${{ always() }}
+        run: echo ${{ github.event.pull_request.number }} > PR_number.txt
+
+      - name: Upload linting log file artifact
+        if: ${{ always() }}
+        uses: actions/upload-artifact@v3
+        with:
+          name: linting-logs
+          path: |
+            lint_log.txt
+            lint_results.md
+            PR_number.txt
diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml
new file mode 100644
index 00000000..0bbcd30f
--- /dev/null
+++ b/.github/workflows/linting_comment.yml
@@ -0,0 +1,28 @@
+name: nf-core linting comment
+# This workflow is triggered after the linting action is complete
+# It posts an automated comment to the PR, even if the PR is coming from a fork
+
+on:
+  workflow_run:
+    workflows: ["nf-core linting"]
+
+jobs:
+  test:
+    runs-on: ubuntu-latest
+    steps:
+      - name: Download lint results
+        uses: dawidd6/action-download-artifact@v2
+        with:
+          workflow: linting.yml
+          workflow_conclusion: completed
+
+      - name: Get PR number
+        id: pr_number
+        run: echo "pr_number=$(cat linting-logs/PR_number.txt)" >> $GITHUB_OUTPUT
+
+      - name: Post PR comment
+        uses: marocchino/sticky-pull-request-comment@v2
+        with:
+          GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }}
+          number: ${{ steps.pr_number.outputs.pr_number }}
+          path: linting-logs/lint_results.md
diff --git a/.gitignore b/.gitignore
new file mode 100644
index 00000000..5124c9ac
--- /dev/null
+++ b/.gitignore
@@ -0,0 +1,8 @@
+.nextflow*
+work/
+data/
+results/
+.DS_Store
+testing/
+testing*
+*.pyc
diff --git a/.gitpod.yml b/.gitpod.yml
new file mode 100644
index 00000000..25488dcc
--- /dev/null
+++ b/.gitpod.yml
@@ -0,0 +1,19 @@
+image: nfcore/gitpod:latest
+tasks:
+  - name: Update Nextflow and setup pre-commit
+    command: |
+      pre-commit install --install-hooks
+      nextflow self-update
+
+vscode:
+  extensions: # based on nf-core.nf-core-extensionpack
+    - codezombiech.gitignore # Language support for .gitignore files
+    # - cssho.vscode-svgviewer                 # SVG viewer
+    - esbenp.prettier-vscode # Markdown/CommonMark linting and style checking for Visual Studio Code
+    - eamodio.gitlens # Quickly glimpse into whom, why, and when a line or code block was changed
+    - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files
+    - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar
+    - mechatroner.rainbow-csv # Highlight columns in csv files in different colors
+    # - nextflow.nextflow                      # Nextflow syntax highlighting
+    - oderwat.indent-rainbow # Highlight indentation level
+    - streetsidesoftware.code-spell-checker # Spelling checker for source code
diff --git a/.nf-core.yml b/.nf-core.yml
new file mode 100644
index 00000000..3805dc81
--- /dev/null
+++ b/.nf-core.yml
@@ -0,0 +1 @@
+repository_type: pipeline
diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml
new file mode 100644
index 00000000..0c31cdb9
--- /dev/null
+++ b/.pre-commit-config.yaml
@@ -0,0 +1,5 @@
+repos:
+  - repo: https://github.com/pre-commit/mirrors-prettier
+    rev: "v2.7.1"
+    hooks:
+      - id: prettier
diff --git a/.prettierignore b/.prettierignore
new file mode 100644
index 00000000..437d763d
--- /dev/null
+++ b/.prettierignore
@@ -0,0 +1,12 @@
+email_template.html
+adaptivecard.json
+slackreport.json
+.nextflow*
+work/
+data/
+results/
+.DS_Store
+testing/
+testing*
+*.pyc
+bin/
diff --git a/.prettierrc.yml b/.prettierrc.yml
new file mode 100644
index 00000000..c81f9a76
--- /dev/null
+++ b/.prettierrc.yml
@@ -0,0 +1 @@
+printWidth: 120
diff --git a/CHANGELOG.md b/CHANGELOG.md
new file mode 100644
index 00000000..686777b5
--- /dev/null
+++ b/CHANGELOG.md
@@ -0,0 +1,16 @@
+# nf-core/rnadeseq: Changelog
+
+The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
+and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
+
+## v2.2dev - [date]
+
+Initial release of nf-core/rnadeseq, created with the [nf-core](https://nf-co.re/) template.
+
+### `Added`
+
+### `Fixed`
+
+### `Dependencies`
+
+### `Deprecated`
diff --git a/CITATIONS.md b/CITATIONS.md
new file mode 100644
index 00000000..124e3fe1
--- /dev/null
+++ b/CITATIONS.md
@@ -0,0 +1,41 @@
+# nf-core/rnadeseq: Citations
+
+## [nf-core](https://pubmed.ncbi.nlm.nih.gov/32055031/)
+
+> Ewels PA, Peltzer A, Fillinger S, Patel H, Alneberg J, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. The nf-core framework for community-curated bioinformatics pipelines. Nat Biotechnol. 2020 Mar;38(3):276-278. doi: 10.1038/s41587-020-0439-x. PubMed PMID: 32055031.
+
+## [Nextflow](https://pubmed.ncbi.nlm.nih.gov/28398311/)
+
+> Di Tommaso P, Chatzou M, Floden EW, Barja PP, Palumbo E, Notredame C. Nextflow enables reproducible computational workflows. Nat Biotechnol. 2017 Apr 11;35(4):316-319. doi: 10.1038/nbt.3820. PubMed PMID: 28398311.
+
+## Pipeline tools
+
+- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)
+
+  > Andrews, S. (2010). FastQC: A Quality Control Tool for High Throughput Sequence Data [Online]. Available online https://www.bioinformatics.babraham.ac.uk/projects/fastqc/.
+
+- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/)
+
+  > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924.
+
+## Software packaging/containerisation tools
+
+- [Anaconda](https://anaconda.com)
+
+  > Anaconda Software Distribution. Computer software. Vers. 2-2.4.0. Anaconda, Nov. 2016. Web.
+
+- [Bioconda](https://pubmed.ncbi.nlm.nih.gov/29967506/)
+
+  > Grüning B, Dale R, Sjödin A, Chapman BA, Rowe J, Tomkins-Tinch CH, Valieris R, Köster J; Bioconda Team. Bioconda: sustainable and comprehensive software distribution for the life sciences. Nat Methods. 2018 Jul;15(7):475-476. doi: 10.1038/s41592-018-0046-7. PubMed PMID: 29967506.
+
+- [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/)
+
+  > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671.
+
+- [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241)
+
+  > Merkel, D. (2014). Docker: lightweight linux containers for consistent development and deployment. Linux Journal, 2014(239), 2. doi: 10.5555/2600239.2600241.
+
+- [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/)
+
+  > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675.
diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md
new file mode 100644
index 00000000..f4fd052f
--- /dev/null
+++ b/CODE_OF_CONDUCT.md
@@ -0,0 +1,111 @@
+# Code of Conduct at nf-core (v1.0)
+
+## Our Pledge
+
+In the interest of fostering an open, collaborative, and welcoming environment, we as contributors and maintainers of nf-core, pledge to making participation in our projects and community a harassment-free experience for everyone, regardless of:
+
+- Age
+- Body size
+- Familial status
+- Gender identity and expression
+- Geographical location
+- Level of experience
+- Nationality and national origins
+- Native language
+- Physical and neurological ability
+- Race or ethnicity
+- Religion
+- Sexual identity and orientation
+- Socioeconomic status
+
+Please note that the list above is alphabetised and is therefore not ranked in any order of preference or importance.
+
+## Preamble
+
+> Note: This Code of Conduct (CoC) has been drafted by the nf-core Safety Officer and been edited after input from members of the nf-core team and others. "We", in this document, refers to the Safety Officer and members of the nf-core core team, both of whom are deemed to be members of the nf-core community and are therefore required to abide by this Code of Conduct. This document will amended periodically to keep it up-to-date, and in case of any dispute, the most current version will apply.
+
+An up-to-date list of members of the nf-core core team can be found [here](https://nf-co.re/about). Our current safety officer is Renuka Kudva.
+
+nf-core is a young and growing community that welcomes contributions from anyone with a shared vision for [Open Science Policies](https://www.fosteropenscience.eu/taxonomy/term/8). Open science policies encompass inclusive behaviours and we strive to build and maintain a safe and inclusive environment for all individuals.
+
+We have therefore adopted this code of conduct (CoC), which we require all members of our community and attendees in nf-core events to adhere to in all our workspaces at all times. Workspaces include but are not limited to Slack, meetings on Zoom, Jitsi, YouTube live etc.
+
+Our CoC will be strictly enforced and the nf-core team reserve the right to exclude participants who do not comply with our guidelines from our workspaces and future nf-core activities.
+
+We ask all members of our community to help maintain a supportive and productive workspace and to avoid behaviours that can make individuals feel unsafe or unwelcome. Please help us maintain and uphold this CoC.
+
+Questions, concerns or ideas on what we can include? Contact safety [at] nf-co [dot] re
+
+## Our Responsibilities
+
+The safety officer is responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behaviour.
+
+The safety officer in consultation with the nf-core core team have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this Code of Conduct, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful.
+
+Members of the core team or the safety officer who violate the CoC will be required to recuse themselves pending investigation. They will not have access to any reports of the violations and be subject to the same actions as others in violation of the CoC.
+
+## When are where does this Code of Conduct apply?
+
+Participation in the nf-core community is contingent on following these guidelines in all our workspaces and events. This includes but is not limited to the following listed alphabetically and therefore in no order of preference:
+
+- Communicating with an official project email address.
+- Communicating with community members within the nf-core Slack channel.
+- Participating in hackathons organised by nf-core (both online and in-person events).
+- Participating in collaborative work on GitHub, Google Suite, community calls, mentorship meetings, email correspondence.
+- Participating in workshops, training, and seminar series organised by nf-core (both online and in-person events). This applies to events hosted on web-based platforms such as Zoom, Jitsi, YouTube live etc.
+- Representing nf-core on social media. This includes both official and personal accounts.
+
+## nf-core cares 😊
+
+nf-core's CoC and expectations of respectful behaviours for all participants (including organisers and the nf-core team) include but are not limited to the following (listed in alphabetical order):
+
+- Ask for consent before sharing another community member’s personal information (including photographs) on social media.
+- Be respectful of differing viewpoints and experiences. We are all here to learn from one another and a difference in opinion can present a good learning opportunity.
+- Celebrate your accomplishments at events! (Get creative with your use of emojis 🎉 🥳 💯 🙌 !)
+- Demonstrate empathy towards other community members. (We don’t all have the same amount of time to dedicate to nf-core. If tasks are pending, don’t hesitate to gently remind members of your team. If you are leading a task, ask for help if you feel overwhelmed.)
+- Engage with and enquire after others. (This is especially important given the geographically remote nature of the nf-core community, so let’s do this the best we can)
+- Focus on what is best for the team and the community. (When in doubt, ask)
+- Graciously accept constructive criticism, yet be unafraid to question, deliberate, and learn.
+- Introduce yourself to members of the community. (We’ve all been outsiders and we know that talking to strangers can be hard for some, but remember we’re interested in getting to know you and your visions for open science!)
+- Show appreciation and **provide clear feedback**. (This is especially important because we don’t see each other in person and it can be harder to interpret subtleties. Also remember that not everyone understands a certain language to the same extent as you do, so **be clear in your communications to be kind.**)
+- Take breaks when you feel like you need them.
+- Using welcoming and inclusive language. (Participants are encouraged to display their chosen pronouns on Zoom or in communication on Slack.)
+
+## nf-core frowns on 😕
+
+The following behaviours from any participants within the nf-core community (including the organisers) will be considered unacceptable under this code of conduct. Engaging or advocating for any of the following could result in expulsion from nf-core workspaces.
+
+- Deliberate intimidation, stalking or following and sustained disruption of communication among participants of the community. This includes hijacking shared screens through actions such as using the annotate tool in conferencing software such as Zoom.
+- “Doxing” i.e. posting (or threatening to post) another person’s personal identifying information online.
+- Spamming or trolling of individuals on social media.
+- Use of sexual or discriminatory imagery, comments, or jokes and unwelcome sexual attention.
+- Verbal and text comments that reinforce social structures of domination related to gender, gender identity and expression, sexual orientation, ability, physical appearance, body size, race, age, religion or work experience.
+
+### Online Trolling
+
+The majority of nf-core interactions and events are held online. Unfortunately, holding events online comes with the added issue of online trolling. This is unacceptable, reports of such behaviour will be taken very seriously, and perpetrators will be excluded from activities immediately.
+
+All community members are required to ask members of the group they are working within for explicit consent prior to taking screenshots of individuals during video calls.
+
+## Procedures for Reporting CoC violations
+
+If someone makes you feel uncomfortable through their behaviours or actions, report it as soon as possible.
+
+You can reach out to members of the [nf-core core team](https://nf-co.re/about) and they will forward your concerns to the safety officer(s).
+
+Issues directly concerning members of the core team will be dealt with by other members of the core team and the safety manager, and possible conflicts of interest will be taken into account. nf-core is also in discussions about having an ombudsperson, and details will be shared in due course.
+
+All reports will be handled with utmost discretion and confidentially.
+
+## Attribution and Acknowledgements
+
+- The [Contributor Covenant, version 1.4](http://contributor-covenant.org/version/1/4)
+- The [OpenCon 2017 Code of Conduct](http://www.opencon2017.org/code_of_conduct) (CC BY 4.0 OpenCon organisers, SPARC and Right to Research Coalition)
+- The [eLife innovation sprint 2020 Code of Conduct](https://sprint.elifesciences.org/code-of-conduct/)
+- The [Mozilla Community Participation Guidelines v3.1](https://www.mozilla.org/en-US/about/governance/policies/participation/) (version 3.1, CC BY-SA 3.0 Mozilla)
+
+## Changelog
+
+### v1.0 - March 12th, 2021
+
+- Complete rewrite from original [Contributor Covenant](http://contributor-covenant.org/) CoC.
diff --git a/LICENSE b/LICENSE
new file mode 100644
index 00000000..9a33bc6e
--- /dev/null
+++ b/LICENSE
@@ -0,0 +1,21 @@
+MIT License
+
+Copyright (c) Gisela Gabernet, Silvia Morini, Oskar Wacker, QBiC
+
+Permission is hereby granted, free of charge, to any person obtaining a copy
+of this software and associated documentation files (the "Software"), to deal
+in the Software without restriction, including without limitation the rights
+to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
+copies of the Software, and to permit persons to whom the Software is
+furnished to do so, subject to the following conditions:
+
+The above copyright notice and this permission notice shall be included in all
+copies or substantial portions of the Software.
+
+THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
+IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
+FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE
+AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
+LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
+OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE
+SOFTWARE.
diff --git a/README.md b/README.md
new file mode 100644
index 00000000..778080eb
--- /dev/null
+++ b/README.md
@@ -0,0 +1,106 @@
+# ![nf-core/rnadeseq](docs/images/nf-core-rnadeseq_logo_light.png#gh-light-mode-only) ![nf-core/rnadeseq](docs/images/nf-core-rnadeseq_logo_dark.png#gh-dark-mode-only)
+
+[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/rnadeseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)
+
+[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.svg)](https://www.nextflow.io/)
+[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)
+[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)
+[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)
+[![Launch on Nextflow Tower](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Nextflow%20Tower-%234256e7)](https://tower.nf/launch?pipeline=https://github.com/nf-core/rnadeseq)
+
+[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23rnadeseq-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/rnadeseq)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)
+
+## Introduction
+
+**nf-core/rnadeseq** is a bioinformatics pipeline that ...
+
+<!-- TODO nf-core:
+   Complete this sentence with a 2-3 sentence summary of what types of data the pipeline ingests, a brief overview of the
+   major pipeline sections and the types of output it produces. You're giving an overview to someone new
+   to nf-core here, in 15-20 seconds. For an example, see https://github.com/nf-core/rnaseq/blob/master/README.md#introduction
+-->
+
+<!-- TODO nf-core: Include a figure that guides the user through the major workflow steps. Many nf-core
+     workflows use the "tube map" design for that. See https://nf-co.re/docs/contributing/design_guidelines#examples for examples.   -->
+<!-- TODO nf-core: Fill in short bullet-pointed list of the default steps in the pipeline -->
+
+1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/))
+2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))
+
+## Usage
+
+> **Note**
+> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how
+> to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline)
+> with `-profile test` before running the workflow on actual data.
+
+<!-- TODO nf-core: Describe the minimum required steps to execute the pipeline, e.g. how to prepare samplesheets.
+     Explain what rows and columns represent. For instance (please edit as appropriate):
+
+First, prepare a samplesheet with your input data that looks as follows:
+
+`samplesheet.csv`:
+
+```csv
+sample,fastq_1,fastq_2
+CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
+```
+
+Each row represents a fastq file (single-end) or a pair of fastq files (paired end).
+
+-->
+
+Now, you can run the pipeline using:
+
+<!-- TODO nf-core: update the following command to include all required parameters for a minimal example -->
+
+```bash
+nextflow run nf-core/rnadeseq \
+   -profile <docker/singularity/.../institute> \
+   --input samplesheet.csv \
+   --outdir <OUTDIR>
+```
+
+> **Warning:**
+> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those
+> provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_;
+> see [docs](https://nf-co.re/usage/configuration#custom-configuration-files).
+
+For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/rnadeseq/usage) and the [parameter documentation](https://nf-co.re/rnadeseq/parameters).
+
+## Pipeline output
+
+To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/rnadeseq/results) tab on the nf-core website pipeline page.
+For more details about the output files and reports, please refer to the
+[output documentation](https://nf-co.re/rnadeseq/output).
+
+## Credits
+
+nf-core/rnadeseq was originally written by Gisela Gabernet, Silvia Morini, Oskar Wacker, QBiC.
+
+We thank the following people for their extensive assistance in the development of this pipeline:
+
+<!-- TODO nf-core: If applicable, make list of people who have also contributed -->
+
+## Contributions and Support
+
+If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md).
+
+For further information or help, don't hesitate to get in touch on the [Slack `#rnadeseq` channel](https://nfcore.slack.com/channels/rnadeseq) (you can join with [this invite](https://nf-co.re/join/slack)).
+
+## Citations
+
+<!-- TODO nf-core: Add citation for pipeline after first release. Uncomment lines below and update Zenodo doi and badge at the top of this file. -->
+<!-- If you use  nf-core/rnadeseq for your analysis, please cite it using the following doi: [10.5281/zenodo.XXXXXX](https://doi.org/10.5281/zenodo.XXXXXX) -->
+
+<!-- TODO nf-core: Add bibliography of tools and data used in your pipeline -->
+
+An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.
+
+You can cite the `nf-core` publication as follows:
+
+> **The nf-core framework for community-curated bioinformatics pipelines.**
+>
+> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.
+>
+> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).
diff --git a/assets/adaptivecard.json b/assets/adaptivecard.json
new file mode 100644
index 00000000..99786880
--- /dev/null
+++ b/assets/adaptivecard.json
@@ -0,0 +1,67 @@
+{
+    "type": "message",
+    "attachments": [
+        {
+            "contentType": "application/vnd.microsoft.card.adaptive",
+            "contentUrl": null,
+            "content": {
+                "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json",
+                "msteams": {
+                    "width": "Full"
+                },
+                "type": "AdaptiveCard",
+                "version": "1.2",
+                "body": [
+                    {
+                        "type": "TextBlock",
+                        "size": "Large",
+                        "weight": "Bolder",
+                        "color": "<% if (success) { %>Good<% } else { %>Attention<%} %>",
+                        "text": "nf-core/rnadeseq v${version} - ${runName}",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "spacing": "None",
+                        "text": "Completed at ${dateComplete} (duration: ${duration})",
+                        "isSubtle": true,
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors. The full error message was: ${errorReport}.<% } %>",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "The command used to launch the workflow was as follows:",
+                        "wrap": true
+                    },
+                    {
+                        "type": "TextBlock",
+                        "text": "${commandLine}",
+                        "isSubtle": true,
+                        "wrap": true
+                    }
+                ],
+                "actions": [
+                    {
+                        "type": "Action.ShowCard",
+                        "title": "Pipeline Configuration",
+                        "card": {
+                            "type": "AdaptiveCard",
+                            "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json",
+                            "body": [
+                                {
+                                    "type": "FactSet",
+                                    "facts": [<% out << summary.collect{ k,v -> "{\"title\": \"$k\", \"value\" : \"$v\"}"}.join(",\n") %>
+                                    ]
+                                }
+                            ]
+                        }
+                    }
+                ]
+            }
+        }
+    ]
+}
diff --git a/assets/email_template.html b/assets/email_template.html
new file mode 100644
index 00000000..174cf877
--- /dev/null
+++ b/assets/email_template.html
@@ -0,0 +1,53 @@
+<html>
+<head>
+  <meta charset="utf-8">
+  <meta http-equiv="X-UA-Compatible" content="IE=edge">
+  <meta name="viewport" content="width=device-width, initial-scale=1">
+
+  <meta name="description" content="nf-core/rnadeseq: Differential gene expression analysis and pathway analysis of RNAseq data">
+  <title>nf-core/rnadeseq Pipeline Report</title>
+</head>
+<body>
+<div style="font-family: Helvetica, Arial, sans-serif; padding: 30px; max-width: 800px; margin: 0 auto;">
+
+<img src="cid:nfcorepipelinelogo">
+
+<h1>nf-core/rnadeseq v${version}</h1>
+<h2>Run Name: $runName</h2>
+
+<% if (!success){
+    out << """
+    <div style="color: #a94442; background-color: #f2dede; border-color: #ebccd1; padding: 15px; margin-bottom: 20px; border: 1px solid transparent; border-radius: 4px;">
+        <h4 style="margin-top:0; color: inherit;">nf-core/rnadeseq execution completed unsuccessfully!</h4>
+        <p>The exit status of the task that caused the workflow execution to fail was: <code>$exitStatus</code>.</p>
+        <p>The full error message was:</p>
+        <pre style="white-space: pre-wrap; overflow: visible; margin-bottom: 0;">${errorReport}</pre>
+    </div>
+    """
+} else {
+    out << """
+    <div style="color: #3c763d; background-color: #dff0d8; border-color: #d6e9c6; padding: 15px; margin-bottom: 20px; border: 1px solid transparent; border-radius: 4px;">
+        nf-core/rnadeseq execution completed successfully!
+    </div>
+    """
+}
+%>
+
+<p>The workflow was completed at <strong>$dateComplete</strong> (duration: <strong>$duration</strong>)</p>
+<p>The command used to launch the workflow was as follows:</p>
+<pre style="white-space: pre-wrap; overflow: visible; background-color: #ededed; padding: 15px; border-radius: 4px; margin-bottom:30px;">$commandLine</pre>
+
+<h3>Pipeline Configuration:</h3>
+<table style="width:100%; max-width:100%; border-spacing: 0; border-collapse: collapse; border:0; margin-bottom: 30px;">
+    <tbody style="border-bottom: 1px solid #ddd;">
+        <% out << summary.collect{ k,v -> "<tr><th style='text-align:left; padding: 8px 0; line-height: 1.42857143; vertical-align: top; border-top: 1px solid #ddd;'>$k</th><td style='text-align:left; padding: 8px; line-height: 1.42857143; vertical-align: top; border-top: 1px solid #ddd;'><pre style='white-space: pre-wrap; overflow: visible;'>$v</pre></td></tr>" }.join("\n") %>
+    </tbody>
+</table>
+
+<p>nf-core/rnadeseq</p>
+<p><a href="https://github.com/nf-core/rnadeseq">https://github.com/nf-core/rnadeseq</a></p>
+
+</div>
+
+</body>
+</html>
diff --git a/assets/email_template.txt b/assets/email_template.txt
new file mode 100644
index 00000000..1a48efa9
--- /dev/null
+++ b/assets/email_template.txt
@@ -0,0 +1,39 @@
+----------------------------------------------------
+                                        ,--./,-.
+        ___     __   __   __   ___     /,-._.--~\\
+  |\\ | |__  __ /  ` /  \\ |__) |__         }  {
+  | \\| |       \\__, \\__/ |  \\ |___     \\`-._,-`-,
+                                        `._,._,'
+  nf-core/rnadeseq v${version}
+----------------------------------------------------
+Run Name: $runName
+
+<% if (success){
+    out << "## nf-core/rnadeseq execution completed successfully! ##"
+} else {
+    out << """####################################################
+## nf-core/rnadeseq execution completed unsuccessfully! ##
+####################################################
+The exit status of the task that caused the workflow execution to fail was: $exitStatus.
+The full error message was:
+
+${errorReport}
+"""
+} %>
+
+
+The workflow was completed at $dateComplete (duration: $duration)
+
+The command used to launch the workflow was as follows:
+
+  $commandLine
+
+
+
+Pipeline Configuration:
+-----------------------
+<% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %>
+
+--
+nf-core/rnadeseq
+https://github.com/nf-core/rnadeseq
diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml
new file mode 100644
index 00000000..04b860b4
--- /dev/null
+++ b/assets/methods_description_template.yml
@@ -0,0 +1,29 @@
+id: "nf-core-rnadeseq-methods-description"
+description: "Suggested text and references to use when describing pipeline usage within the methods section of a publication."
+section_name: "nf-core/rnadeseq Methods Description"
+section_href: "https://github.com/nf-core/rnadeseq"
+plot_type: "html"
+## TODO nf-core: Update the HTML below to your preferred methods description, e.g. add publication citation for this pipeline
+## You inject any metadata in the Nextflow '${workflow}' object
+data: |
+  <h4>Methods</h4>
+  <p>Data was processed using nf-core/rnadeseq v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (<a href="https://doi.org/10.1038/s41587-020-0439-x">Ewels <em>et al.</em>, 2020</a>), utilising reproducible software environments from the Bioconda (<a href="https://doi.org/10.1038/s41592-018-0046-7">Grüning <em>et al.</em>, 2018</a>) and Biocontainers (<a href="https://doi.org/10.1093/bioinformatics/btx192">da Veiga Leprevost <em>et al.</em>, 2017</a>) projects.</p>
+  <p>The pipeline was executed with Nextflow v${workflow.nextflow.version} (<a href="https://doi.org/10.1038/nbt.3820">Di Tommaso <em>et al.</em>, 2017</a>) with the following command:</p>
+  <pre><code>${workflow.commandLine}</code></pre>
+  <p>${tool_citations}</p>
+  <h4>References</h4>
+  <ul>
+    <li>Di Tommaso, P., Chatzou, M., Floden, E. W., Barja, P. P., Palumbo, E., & Notredame, C. (2017). Nextflow enables reproducible computational workflows. Nature Biotechnology, 35(4), 316-319. doi: <a href="https://doi.org/10.1038/nbt.3820">10.1038/nbt.3820</a></li>
+    <li>Ewels, P. A., Peltzer, A., Fillinger, S., Patel, H., Alneberg, J., Wilm, A., Garcia, M. U., Di Tommaso, P., & Nahnsen, S. (2020). The nf-core framework for community-curated bioinformatics pipelines. Nature Biotechnology, 38(3), 276-278. doi: <a href="https://doi.org/10.1038/s41587-020-0439-x">10.1038/s41587-020-0439-x</a></li>
+    <li>Grüning, B., Dale, R., Sjödin, A., Chapman, B. A., Rowe, J., Tomkins-Tinch, C. H., Valieris, R., Köster, J., & Bioconda Team. (2018). Bioconda: sustainable and comprehensive software distribution for the life sciences. Nature Methods, 15(7), 475–476. doi: <a href="https://doi.org/10.1038/s41592-018-0046-7">10.1038/s41592-018-0046-7</a></li>
+    <li>da Veiga Leprevost, F., Grüning, B. A., Alves Aflitos, S., Röst, H. L., Uszkoreit, J., Barsnes, H., Vaudel, M., Moreno, P., Gatto, L., Weber, J., Bai, M., Jimenez, R. C., Sachsenberg, T., Pfeuffer, J., Vera Alvarez, R., Griss, J., Nesvizhskii, A. I., & Perez-Riverol, Y. (2017). BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics (Oxford, England), 33(16), 2580–2582. doi: <a href="https://doi.org/10.1093/bioinformatics/btx192">10.1093/bioinformatics/btx192</a></li>
+    ${tool_bibliography}
+  </ul>
+  <div class="alert alert-info">
+    <h5>Notes:</h5>
+    <ul>
+      ${nodoi_text}
+      <li>The command above does not include parameters contained in any configs or profiles that may have been used. Ensure the config file is also uploaded with your publication!</li>
+      <li>You should also cite all software used within this run. Check the "Software Versions" of this report to get version information.</li>
+    </ul>
+  </div>
diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
new file mode 100644
index 00000000..68bd73a1
--- /dev/null
+++ b/assets/multiqc_config.yml
@@ -0,0 +1,13 @@
+report_comment: >
+  This report has been generated by the <a href="https://github.com/nf-core/rnadeseq/2.2dev" target="_blank">nf-core/rnadeseq</a>
+  analysis pipeline. For information about how to interpret these results, please see the
+  <a href="https://nf-co.re/rnadeseq/2.2dev/output" target="_blank">documentation</a>.
+report_section_order:
+  "nf-core-rnadeseq-methods-description":
+    order: -1000
+  software_versions:
+    order: -1001
+  "nf-core-rnadeseq-summary":
+    order: -1002
+
+export_plots: true
diff --git a/assets/nf-core-rnadeseq_logo_light.png b/assets/nf-core-rnadeseq_logo_light.png
new file mode 100644
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HcmV?d00001

diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv
new file mode 100644
index 00000000..5f653ab7
--- /dev/null
+++ b/assets/samplesheet.csv
@@ -0,0 +1,3 @@
+sample,fastq_1,fastq_2
+SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz
+SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz,
diff --git a/assets/schema_input.json b/assets/schema_input.json
new file mode 100644
index 00000000..bd54c145
--- /dev/null
+++ b/assets/schema_input.json
@@ -0,0 +1,36 @@
+{
+    "$schema": "http://json-schema.org/draft-07/schema",
+    "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/assets/schema_input.json",
+    "title": "nf-core/rnadeseq pipeline - params.input schema",
+    "description": "Schema for the file provided with params.input",
+    "type": "array",
+    "items": {
+        "type": "object",
+        "properties": {
+            "sample": {
+                "type": "string",
+                "pattern": "^\\S+$",
+                "errorMessage": "Sample name must be provided and cannot contain spaces"
+            },
+            "fastq_1": {
+                "type": "string",
+                "pattern": "^\\S+\\.f(ast)?q\\.gz$",
+                "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'"
+            },
+            "fastq_2": {
+                "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'",
+                "anyOf": [
+                    {
+                        "type": "string",
+                        "pattern": "^\\S+\\.f(ast)?q\\.gz$"
+                    },
+                    {
+                        "type": "string",
+                        "maxLength": 0
+                    }
+                ]
+            }
+        },
+        "required": ["sample", "fastq_1"]
+    }
+}
diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt
new file mode 100644
index 00000000..953c8470
--- /dev/null
+++ b/assets/sendmail_template.txt
@@ -0,0 +1,53 @@
+To: $email
+Subject: $subject
+Mime-Version: 1.0
+Content-Type: multipart/related;boundary="nfcoremimeboundary"
+
+--nfcoremimeboundary
+Content-Type: text/html; charset=utf-8
+
+$email_html
+
+--nfcoremimeboundary
+Content-Type: image/png;name="nf-core-rnadeseq_logo.png"
+Content-Transfer-Encoding: base64
+Content-ID: <nfcorepipelinelogo>
+Content-Disposition: inline; filename="nf-core-rnadeseq_logo_light.png"
+
+<% out << new File("$projectDir/assets/nf-core-rnadeseq_logo_light.png").
+    bytes.
+    encodeBase64().
+    toString().
+    tokenize( '\n' )*.
+    toList()*.
+    collate( 76 )*.
+    collect { it.join() }.
+    flatten().
+    join( '\n' ) %>
+
+<%
+if (mqcFile){
+def mqcFileObj = new File("$mqcFile")
+if (mqcFileObj.length() < mqcMaxSize){
+out << """
+--nfcoremimeboundary
+Content-Type: text/html; name=\"multiqc_report\"
+Content-Transfer-Encoding: base64
+Content-ID: <mqcreport>
+Content-Disposition: attachment; filename=\"${mqcFileObj.getName()}\"
+
+${mqcFileObj.
+    bytes.
+    encodeBase64().
+    toString().
+    tokenize( '\n' )*.
+    toList()*.
+    collate( 76 )*.
+    collect { it.join() }.
+    flatten().
+    join( '\n' )}
+"""
+}}
+%>
+
+--nfcoremimeboundary--
diff --git a/assets/slackreport.json b/assets/slackreport.json
new file mode 100644
index 00000000..6f0c493b
--- /dev/null
+++ b/assets/slackreport.json
@@ -0,0 +1,34 @@
+{
+    "attachments": [
+        {
+            "fallback": "Plain-text summary of the attachment.",
+            "color": "<% if (success) { %>good<% } else { %>danger<%} %>",
+            "author_name": "nf-core/rnadeseq v${version} - ${runName}",
+            "author_icon": "https://www.nextflow.io/docs/latest/_static/favicon.ico",
+            "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors<% } %>",
+            "fields": [
+                {
+                    "title": "Command used to launch the workflow",
+                    "value": "```${commandLine}```",
+                    "short": false
+                }
+                <%
+                    if (!success) { %>
+                    ,
+                    {
+                        "title": "Full error message",
+                        "value": "```${errorReport}```",
+                        "short": false
+                    },
+                    {
+                        "title": "Pipeline configuration",
+                        "value": "<% out << summary.collect{ k,v -> k == "hook_url" ? "_${k}_: (_hidden_)" : ( ( v.class.toString().contains('Path') || ( v.class.toString().contains('String') && v.contains('/') ) ) ? "_${k}_: `${v}`" : (v.class.toString().contains('DateTime') ? ("_${k}_: " + v.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM))) : "_${k}_: ${v}") ) }.join(",\n") %>",
+                        "short": false
+                    }
+                    <% }
+                %>
+            ],
+            "footer": "Completed at <% out << dateComplete.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM)) %> (duration: ${duration})"
+        }
+    ]
+}
diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py
new file mode 100755
index 00000000..4a758fe0
--- /dev/null
+++ b/bin/check_samplesheet.py
@@ -0,0 +1,259 @@
+#!/usr/bin/env python
+
+
+"""Provide a command line tool to validate and transform tabular samplesheets."""
+
+
+import argparse
+import csv
+import logging
+import sys
+from collections import Counter
+from pathlib import Path
+
+logger = logging.getLogger()
+
+
+class RowChecker:
+    """
+    Define a service that can validate and transform each given row.
+
+    Attributes:
+        modified (list): A list of dicts, where each dict corresponds to a previously
+            validated and transformed row. The order of rows is maintained.
+
+    """
+
+    VALID_FORMATS = (
+        ".fq.gz",
+        ".fastq.gz",
+    )
+
+    def __init__(
+        self,
+        sample_col="sample",
+        first_col="fastq_1",
+        second_col="fastq_2",
+        single_col="single_end",
+        **kwargs,
+    ):
+        """
+        Initialize the row checker with the expected column names.
+
+        Args:
+            sample_col (str): The name of the column that contains the sample name
+                (default "sample").
+            first_col (str): The name of the column that contains the first (or only)
+                FASTQ file path (default "fastq_1").
+            second_col (str): The name of the column that contains the second (if any)
+                FASTQ file path (default "fastq_2").
+            single_col (str): The name of the new column that will be inserted and
+                records whether the sample contains single- or paired-end sequencing
+                reads (default "single_end").
+
+        """
+        super().__init__(**kwargs)
+        self._sample_col = sample_col
+        self._first_col = first_col
+        self._second_col = second_col
+        self._single_col = single_col
+        self._seen = set()
+        self.modified = []
+
+    def validate_and_transform(self, row):
+        """
+        Perform all validations on the given row and insert the read pairing status.
+
+        Args:
+            row (dict): A mapping from column headers (keys) to elements of that row
+                (values).
+
+        """
+        self._validate_sample(row)
+        self._validate_first(row)
+        self._validate_second(row)
+        self._validate_pair(row)
+        self._seen.add((row[self._sample_col], row[self._first_col]))
+        self.modified.append(row)
+
+    def _validate_sample(self, row):
+        """Assert that the sample name exists and convert spaces to underscores."""
+        if len(row[self._sample_col]) <= 0:
+            raise AssertionError("Sample input is required.")
+        # Sanitize samples slightly.
+        row[self._sample_col] = row[self._sample_col].replace(" ", "_")
+
+    def _validate_first(self, row):
+        """Assert that the first FASTQ entry is non-empty and has the right format."""
+        if len(row[self._first_col]) <= 0:
+            raise AssertionError("At least the first FASTQ file is required.")
+        self._validate_fastq_format(row[self._first_col])
+
+    def _validate_second(self, row):
+        """Assert that the second FASTQ entry has the right format if it exists."""
+        if len(row[self._second_col]) > 0:
+            self._validate_fastq_format(row[self._second_col])
+
+    def _validate_pair(self, row):
+        """Assert that read pairs have the same file extension. Report pair status."""
+        if row[self._first_col] and row[self._second_col]:
+            row[self._single_col] = False
+            first_col_suffix = Path(row[self._first_col]).suffixes[-2:]
+            second_col_suffix = Path(row[self._second_col]).suffixes[-2:]
+            if first_col_suffix != second_col_suffix:
+                raise AssertionError("FASTQ pairs must have the same file extensions.")
+        else:
+            row[self._single_col] = True
+
+    def _validate_fastq_format(self, filename):
+        """Assert that a given filename has one of the expected FASTQ extensions."""
+        if not any(filename.endswith(extension) for extension in self.VALID_FORMATS):
+            raise AssertionError(
+                f"The FASTQ file has an unrecognized extension: {filename}\n"
+                f"It should be one of: {', '.join(self.VALID_FORMATS)}"
+            )
+
+    def validate_unique_samples(self):
+        """
+        Assert that the combination of sample name and FASTQ filename is unique.
+
+        In addition to the validation, also rename all samples to have a suffix of _T{n}, where n is the
+        number of times the same sample exist, but with different FASTQ files, e.g., multiple runs per experiment.
+
+        """
+        if len(self._seen) != len(self.modified):
+            raise AssertionError("The pair of sample name and FASTQ must be unique.")
+        seen = Counter()
+        for row in self.modified:
+            sample = row[self._sample_col]
+            seen[sample] += 1
+            row[self._sample_col] = f"{sample}_T{seen[sample]}"
+
+
+def read_head(handle, num_lines=10):
+    """Read the specified number of lines from the current position in the file."""
+    lines = []
+    for idx, line in enumerate(handle):
+        if idx == num_lines:
+            break
+        lines.append(line)
+    return "".join(lines)
+
+
+def sniff_format(handle):
+    """
+    Detect the tabular format.
+
+    Args:
+        handle (text file): A handle to a `text file`_ object. The read position is
+        expected to be at the beginning (index 0).
+
+    Returns:
+        csv.Dialect: The detected tabular format.
+
+    .. _text file:
+        https://docs.python.org/3/glossary.html#term-text-file
+
+    """
+    peek = read_head(handle)
+    handle.seek(0)
+    sniffer = csv.Sniffer()
+    dialect = sniffer.sniff(peek)
+    return dialect
+
+
+def check_samplesheet(file_in, file_out):
+    """
+    Check that the tabular samplesheet has the structure expected by nf-core pipelines.
+
+    Validate the general shape of the table, expected columns, and each row. Also add
+    an additional column which records whether one or two FASTQ reads were found.
+
+    Args:
+        file_in (pathlib.Path): The given tabular samplesheet. The format can be either
+            CSV, TSV, or any other format automatically recognized by ``csv.Sniffer``.
+        file_out (pathlib.Path): Where the validated and transformed samplesheet should
+            be created; always in CSV format.
+
+    Example:
+        This function checks that the samplesheet follows the following structure,
+        see also the `viral recon samplesheet`_::
+
+            sample,fastq_1,fastq_2
+            SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz
+            SAMPLE_PE,SAMPLE_PE_RUN2_1.fastq.gz,SAMPLE_PE_RUN2_2.fastq.gz
+            SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz,
+
+    .. _viral recon samplesheet:
+        https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv
+
+    """
+    required_columns = {"sample", "fastq_1", "fastq_2"}
+    # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`.
+    with file_in.open(newline="") as in_handle:
+        reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle))
+        # Validate the existence of the expected header columns.
+        if not required_columns.issubset(reader.fieldnames):
+            req_cols = ", ".join(required_columns)
+            logger.critical(f"The sample sheet **must** contain these column headers: {req_cols}.")
+            sys.exit(1)
+        # Validate each row.
+        checker = RowChecker()
+        for i, row in enumerate(reader):
+            try:
+                checker.validate_and_transform(row)
+            except AssertionError as error:
+                logger.critical(f"{str(error)} On line {i + 2}.")
+                sys.exit(1)
+        checker.validate_unique_samples()
+    header = list(reader.fieldnames)
+    header.insert(1, "single_end")
+    # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`.
+    with file_out.open(mode="w", newline="") as out_handle:
+        writer = csv.DictWriter(out_handle, header, delimiter=",")
+        writer.writeheader()
+        for row in checker.modified:
+            writer.writerow(row)
+
+
+def parse_args(argv=None):
+    """Define and immediately parse command line arguments."""
+    parser = argparse.ArgumentParser(
+        description="Validate and transform a tabular samplesheet.",
+        epilog="Example: python check_samplesheet.py samplesheet.csv samplesheet.valid.csv",
+    )
+    parser.add_argument(
+        "file_in",
+        metavar="FILE_IN",
+        type=Path,
+        help="Tabular input samplesheet in CSV or TSV format.",
+    )
+    parser.add_argument(
+        "file_out",
+        metavar="FILE_OUT",
+        type=Path,
+        help="Transformed output samplesheet in CSV format.",
+    )
+    parser.add_argument(
+        "-l",
+        "--log-level",
+        help="The desired log level (default WARNING).",
+        choices=("CRITICAL", "ERROR", "WARNING", "INFO", "DEBUG"),
+        default="WARNING",
+    )
+    return parser.parse_args(argv)
+
+
+def main(argv=None):
+    """Coordinate argument parsing and program execution."""
+    args = parse_args(argv)
+    logging.basicConfig(level=args.log_level, format="[%(levelname)s] %(message)s")
+    if not args.file_in.is_file():
+        logger.error(f"The given input file {args.file_in} was not found!")
+        sys.exit(2)
+    args.file_out.parent.mkdir(parents=True, exist_ok=True)
+    check_samplesheet(args.file_in, args.file_out)
+
+
+if __name__ == "__main__":
+    sys.exit(main())
diff --git a/conf/base.config b/conf/base.config
new file mode 100644
index 00000000..a5194811
--- /dev/null
+++ b/conf/base.config
@@ -0,0 +1,65 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq Nextflow base config file
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    A 'blank slate' config file, appropriate for general use on most high performance
+    compute environments. Assumes that all software is installed and available on
+    the PATH. Runs in `local` mode - all jobs will be run on the logged in environment.
+----------------------------------------------------------------------------------------
+*/
+
+process {
+
+    // TODO nf-core: Check the defaults for all processes
+    cpus   = { check_max( 1    * task.attempt, 'cpus'   ) }
+    memory = { check_max( 6.GB * task.attempt, 'memory' ) }
+    time   = { check_max( 4.h  * task.attempt, 'time'   ) }
+
+    errorStrategy = { task.exitStatus in ((130..145) + 104) ? 'retry' : 'finish' }
+    maxRetries    = 1
+    maxErrors     = '-1'
+
+    // Process-specific resource requirements
+    // NOTE - Please try and re-use the labels below as much as possible.
+    //        These labels are used and recognised by default in DSL2 files hosted on nf-core/modules.
+    //        If possible, it would be nice to keep the same label naming convention when
+    //        adding in your local modules too.
+    // TODO nf-core: Customise requirements for specific processes.
+    // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors
+    withLabel:process_single {
+        cpus   = { check_max( 1                  , 'cpus'    ) }
+        memory = { check_max( 6.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 4.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_low {
+        cpus   = { check_max( 2     * task.attempt, 'cpus'    ) }
+        memory = { check_max( 12.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 4.h   * task.attempt, 'time'    ) }
+    }
+    withLabel:process_medium {
+        cpus   = { check_max( 6     * task.attempt, 'cpus'    ) }
+        memory = { check_max( 36.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 8.h   * task.attempt, 'time'    ) }
+    }
+    withLabel:process_high {
+        cpus   = { check_max( 12    * task.attempt, 'cpus'    ) }
+        memory = { check_max( 72.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 16.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_long {
+        time   = { check_max( 20.h  * task.attempt, 'time'    ) }
+    }
+    withLabel:process_high_memory {
+        memory = { check_max( 200.GB * task.attempt, 'memory' ) }
+    }
+    withLabel:error_ignore {
+        errorStrategy = 'ignore'
+    }
+    withLabel:error_retry {
+        errorStrategy = 'retry'
+        maxRetries    = 2
+    }
+    withName:CUSTOM_DUMPSOFTWAREVERSIONS {
+        cache = false
+    }
+}
diff --git a/conf/igenomes.config b/conf/igenomes.config
new file mode 100644
index 00000000..3f114377
--- /dev/null
+++ b/conf/igenomes.config
@@ -0,0 +1,440 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for iGenomes paths
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines reference genomes using iGenome paths.
+    Can be used by any config that customises the base path using:
+        $params.igenomes_base / --igenomes_base
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    // illumina iGenomes reference file paths
+    genomes {
+        'GRCh37' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/GRCh37-blacklist.bed"
+        }
+        'GRCh38' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg38-blacklist.bed"
+        }
+        'CHM13' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/BWAIndex/"
+            bwamem2     = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/BWAmem2Index/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/NCBI/CHM13/Annotation/Genes/genes.gtf"
+            gff         = "ftp://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/009/914/755/GCF_009914755.1_T2T-CHM13v2.0/GCF_009914755.1_T2T-CHM13v2.0_genomic.gff.gz"
+            mito_name   = "chrM"
+        }
+        'GRCm38' {
+            fasta       = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "1.87e9"
+            blacklist   = "${projectDir}/assets/blacklists/GRCm38-blacklist.bed"
+        }
+        'TAIR10' {
+            fasta       = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/README.txt"
+            mito_name   = "Mt"
+        }
+        'EB2' {
+            fasta       = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/README.txt"
+        }
+        'UMD3.1' {
+            fasta       = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'WBcel235' {
+            fasta       = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed"
+            mito_name   = "MtDNA"
+            macs_gsize  = "9e7"
+        }
+        'CanFam3.1' {
+            fasta       = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'GRCz10' {
+            fasta       = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'BDGP6' {
+            fasta       = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed"
+            mito_name   = "M"
+            macs_gsize  = "1.2e8"
+        }
+        'EquCab2' {
+            fasta       = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'EB1' {
+            fasta       = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/README.txt"
+        }
+        'Galgal4' {
+            fasta       = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'Gm01' {
+            fasta       = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/README.txt"
+        }
+        'Mmul_1' {
+            fasta       = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'IRGSP-1.0' {
+            fasta       = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed"
+            mito_name   = "Mt"
+        }
+        'CHIMP2.1.4' {
+            fasta       = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'Rnor_5.0' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'Rnor_6.0' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+        }
+        'R64-1-1' {
+            fasta       = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed"
+            mito_name   = "MT"
+            macs_gsize  = "1.2e7"
+        }
+        'EF2' {
+            fasta       = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/README.txt"
+            mito_name   = "MT"
+            macs_gsize  = "1.21e7"
+        }
+        'Sbi1' {
+            fasta       = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/README.txt"
+        }
+        'Sscrofa10.2' {
+            fasta       = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/README.txt"
+            mito_name   = "MT"
+        }
+        'AGPv3' {
+            fasta       = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed"
+            mito_name   = "Mt"
+        }
+        'hg38' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg38-blacklist.bed"
+        }
+        'hg19' {
+            fasta       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "2.7e9"
+            blacklist   = "${projectDir}/assets/blacklists/hg19-blacklist.bed"
+        }
+        'mm10' {
+            fasta       = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "1.87e9"
+            blacklist   = "${projectDir}/assets/blacklists/mm10-blacklist.bed"
+        }
+        'bosTau8' {
+            fasta       = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+        }
+        'ce10' {
+            fasta       = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "9e7"
+        }
+        'canFam3' {
+            fasta       = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'danRer10' {
+            fasta       = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "1.37e9"
+        }
+        'dm6' {
+            fasta       = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+            macs_gsize  = "1.2e8"
+        }
+        'equCab2' {
+            fasta       = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'galGal4' {
+            fasta       = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'panTro4' {
+            fasta       = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+        'rn6' {
+            fasta       = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.bed"
+            mito_name   = "chrM"
+        }
+        'sacCer3' {
+            fasta       = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/"
+            readme      = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Annotation/README.txt"
+            mito_name   = "chrM"
+            macs_gsize  = "1.2e7"
+        }
+        'susScr3' {
+            fasta       = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa"
+            bwa         = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/version0.6.0/"
+            bowtie2     = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/"
+            star        = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/"
+            bismark     = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/"
+            gtf         = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf"
+            bed12       = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed"
+            readme      = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/README.txt"
+            mito_name   = "chrM"
+        }
+    }
+}
diff --git a/conf/modules.config b/conf/modules.config
new file mode 100644
index 00000000..da58a5d8
--- /dev/null
+++ b/conf/modules.config
@@ -0,0 +1,41 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Config file for defining DSL2 per module options and publishing paths
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Available keys to override module options:
+        ext.args   = Additional arguments appended to command in module.
+        ext.args2  = Second set of arguments appended to command in module (multi-tool modules).
+        ext.args3  = Third set of arguments appended to command in module (multi-tool modules).
+        ext.prefix = File name prefix for output files.
+----------------------------------------------------------------------------------------
+*/
+
+process {
+
+    publishDir = [
+        path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" },
+        mode: params.publish_dir_mode,
+        saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+    ]
+
+    withName: SAMPLESHEET_CHECK {
+        publishDir = [
+            path: { "${params.outdir}/pipeline_info" },
+            mode: params.publish_dir_mode,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: FASTQC {
+        ext.args = '--quiet'
+    }
+
+    withName: CUSTOM_DUMPSOFTWAREVERSIONS {
+        publishDir = [
+            path: { "${params.outdir}/pipeline_info" },
+            mode: params.publish_dir_mode,
+            pattern: '*_versions.yml'
+        ]
+    }
+
+}
diff --git a/conf/test.config b/conf/test.config
new file mode 100644
index 00000000..918ce810
--- /dev/null
+++ b/conf/test.config
@@ -0,0 +1,29 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for running minimal tests
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines input files and everything required to run a fast and simple pipeline test.
+
+    Use as follows:
+        nextflow run nf-core/rnadeseq -profile test,<docker/singularity> --outdir <OUTDIR>
+
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    config_profile_name        = 'Test profile'
+    config_profile_description = 'Minimal test dataset to check pipeline function'
+
+    // Limit resources so that this can run on GitHub Actions
+    max_cpus   = 2
+    max_memory = '6.GB'
+    max_time   = '6.h'
+
+    // Input data
+    // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets
+    // TODO nf-core: Give any required params for the test so that command line flags are not needed
+    input  = 'https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv'
+
+    // Genome references
+    genome = 'R64-1-1'
+}
diff --git a/conf/test_full.config b/conf/test_full.config
new file mode 100644
index 00000000..126f6d5a
--- /dev/null
+++ b/conf/test_full.config
@@ -0,0 +1,24 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Nextflow config file for running full-size tests
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Defines input files and everything required to run a full size pipeline test.
+
+    Use as follows:
+        nextflow run nf-core/rnadeseq -profile test_full,<docker/singularity> --outdir <OUTDIR>
+
+----------------------------------------------------------------------------------------
+*/
+
+params {
+    config_profile_name        = 'Full test profile'
+    config_profile_description = 'Full test dataset to check pipeline function'
+
+    // Input data for full size test
+    // TODO nf-core: Specify the paths to your full test data ( on nf-core/test-datasets or directly in repositories, e.g. SRA)
+    // TODO nf-core: Give any required params for the test so that command line flags are not needed
+    input = 'https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_full_illumina_amplicon.csv'
+
+    // Genome references
+    genome = 'R64-1-1'
+}
diff --git a/docs/README.md b/docs/README.md
new file mode 100644
index 00000000..dc052cc1
--- /dev/null
+++ b/docs/README.md
@@ -0,0 +1,10 @@
+# nf-core/rnadeseq: Documentation
+
+The nf-core/rnadeseq documentation is split into the following pages:
+
+- [Usage](usage.md)
+  - An overview of how the pipeline works, how to run it and a description of all of the different command-line flags.
+- [Output](output.md)
+  - An overview of the different results produced by the pipeline and how to interpret them.
+
+You can find a lot more documentation about installing, configuring and running nf-core pipelines on the website: [https://nf-co.re](https://nf-co.re)
diff --git a/docs/images/mqc_fastqc_adapter.png b/docs/images/mqc_fastqc_adapter.png
new file mode 100755
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diff --git a/docs/images/mqc_fastqc_quality.png b/docs/images/mqc_fastqc_quality.png
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diff --git a/docs/images/nf-core-rnadeseq_logo_dark.png b/docs/images/nf-core-rnadeseq_logo_dark.png
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diff --git a/docs/output.md b/docs/output.md
new file mode 100644
index 00000000..34c44ec1
--- /dev/null
+++ b/docs/output.md
@@ -0,0 +1,68 @@
+# nf-core/rnadeseq: Output
+
+## Introduction
+
+This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline.
+
+The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory.
+
+<!-- TODO nf-core: Write this documentation describing your workflow's output -->
+
+## Pipeline overview
+
+The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps:
+
+- [FastQC](#fastqc) - Raw read QC
+- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline
+- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution
+
+### FastQC
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `fastqc/`
+  - `*_fastqc.html`: FastQC report containing quality metrics.
+  - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images.
+
+</details>
+
+[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your sequenced reads. It provides information about the quality score distribution across your reads, per base sequence content (%A/T/G/C), adapter contamination and overrepresented sequences. For further reading and documentation see the [FastQC help pages](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/).
+
+![MultiQC - FastQC sequence counts plot](images/mqc_fastqc_counts.png)
+
+![MultiQC - FastQC mean quality scores plot](images/mqc_fastqc_quality.png)
+
+![MultiQC - FastQC adapter content plot](images/mqc_fastqc_adapter.png)
+
+> **NB:** The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality.
+
+### MultiQC
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `multiqc/`
+  - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
+  - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
+  - `multiqc_plots/`: directory containing static images from the report in various formats.
+
+</details>
+
+[MultiQC](http://multiqc.info) is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory.
+
+Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see <http://multiqc.info>.
+
+### Pipeline information
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `pipeline_info/`
+  - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`.
+  - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline.
+  - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`.
+
+</details>
+
+[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to troubleshoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage.
diff --git a/docs/usage.md b/docs/usage.md
new file mode 100644
index 00000000..44f2647c
--- /dev/null
+++ b/docs/usage.md
@@ -0,0 +1,216 @@
+# nf-core/rnadeseq: Usage
+
+## :warning: Please read this documentation on the nf-core website: [https://nf-co.re/rnadeseq/usage](https://nf-co.re/rnadeseq/usage)
+
+> _Documentation of pipeline parameters is generated automatically from the pipeline schema and can no longer be found in markdown files._
+
+## Introduction
+
+<!-- TODO nf-core: Add documentation about anything specific to running your pipeline. For general topics, please point to (and add to) the main nf-core website. -->
+
+## Samplesheet input
+
+You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below.
+
+```bash
+--input '[path to samplesheet file]'
+```
+
+### Multiple runs of the same sample
+
+The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. The pipeline will concatenate the raw reads before performing any downstream analysis. Below is an example for the same sample sequenced across 3 lanes:
+
+```console
+sample,fastq_1,fastq_2
+CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
+CONTROL_REP1,AEG588A1_S1_L003_R1_001.fastq.gz,AEG588A1_S1_L003_R2_001.fastq.gz
+CONTROL_REP1,AEG588A1_S1_L004_R1_001.fastq.gz,AEG588A1_S1_L004_R2_001.fastq.gz
+```
+
+### Full samplesheet
+
+The pipeline will auto-detect whether a sample is single- or paired-end using the information provided in the samplesheet. The samplesheet can have as many columns as you desire, however, there is a strict requirement for the first 3 columns to match those defined in the table below.
+
+A final samplesheet file consisting of both single- and paired-end data may look something like the one below. This is for 6 samples, where `TREATMENT_REP3` has been sequenced twice.
+
+```console
+sample,fastq_1,fastq_2
+CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
+CONTROL_REP2,AEG588A2_S2_L002_R1_001.fastq.gz,AEG588A2_S2_L002_R2_001.fastq.gz
+CONTROL_REP3,AEG588A3_S3_L002_R1_001.fastq.gz,AEG588A3_S3_L002_R2_001.fastq.gz
+TREATMENT_REP1,AEG588A4_S4_L003_R1_001.fastq.gz,
+TREATMENT_REP2,AEG588A5_S5_L003_R1_001.fastq.gz,
+TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz,
+TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz,
+```
+
+| Column    | Description                                                                                                                                                                            |
+| --------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
+| `sample`  | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). |
+| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz".                                                             |
+| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz".                                                             |
+
+An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline.
+
+## Running the pipeline
+
+The typical command for running the pipeline is as follows:
+
+```bash
+nextflow run nf-core/rnadeseq --input ./samplesheet.csv --outdir ./results --genome GRCh37 -profile docker
+```
+
+This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles.
+
+Note that the pipeline will create the following files in your working directory:
+
+```bash
+work                # Directory containing the nextflow working files
+<OUTDIR>            # Finished results in specified location (defined with --outdir)
+.nextflow_log       # Log file from Nextflow
+# Other nextflow hidden files, eg. history of pipeline runs and old logs.
+```
+
+If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file.
+
+Pipeline settings can be provided in a `yaml` or `json` file via `-params-file <file>`.
+
+> ⚠️ Do not use `-c <file>` to specify parameters as this will result in errors. Custom config files specified with `-c` must only be used for [tuning process resource specifications](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources), other infrastructural tweaks (such as output directories), or module arguments (args).
+
+The above pipeline run specified with a params file in yaml format:
+
+```bash
+nextflow run nf-core/rnadeseq -profile docker -params-file params.yaml
+```
+
+with `params.yaml` containing:
+
+```yaml
+input: './samplesheet.csv'
+outdir: './results/'
+genome: 'GRCh37'
+<...>
+```
+
+You can also generate such `YAML`/`JSON` files via [nf-core/launch](https://nf-co.re/launch).
+
+### Updating the pipeline
+
+When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline:
+
+```bash
+nextflow pull nf-core/rnadeseq
+```
+
+### Reproducibility
+
+It is a good idea to specify a pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since.
+
+First, go to the [nf-core/rnadeseq releases page](https://github.com/nf-core/rnadeseq/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag.
+
+This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. For example, at the bottom of the MultiQC reports.
+
+To further assist in reproducbility, you can use share and re-use [parameter files](#running-the-pipeline) to repeat pipeline runs with the same settings without having to write out a command with every single parameter.
+
+> 💡 If you wish to share such profile (such as upload as supplementary material for academic publications), make sure to NOT include cluster specific paths to files, nor institutional specific profiles.
+
+## Core Nextflow arguments
+
+> **NB:** These options are part of Nextflow and use a _single_ hyphen (pipeline parameters use a double-hyphen).
+
+### `-profile`
+
+Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments.
+
+Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Podman, Shifter, Charliecloud, Apptainer, Conda) - see below.
+
+> We highly recommend the use of Docker or Singularity containers for full pipeline reproducibility, however when this is not possible, Conda is also supported.
+
+The pipeline also dynamically loads configurations from [https://github.com/nf-core/configs](https://github.com/nf-core/configs) when it runs, making multiple config profiles for various institutional clusters available at run time. For more information and to see if your system is available in these configs please see the [nf-core/configs documentation](https://github.com/nf-core/configs#documentation).
+
+Note that multiple profiles can be loaded, for example: `-profile test,docker` - the order of arguments is important!
+They are loaded in sequence, so later profiles can overwrite earlier profiles.
+
+If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended, since it can lead to different results on different machines dependent on the computer enviroment.
+
+- `test`
+  - A profile with a complete configuration for automated testing
+  - Includes links to test data so needs no other parameters
+- `docker`
+  - A generic configuration profile to be used with [Docker](https://docker.com/)
+- `singularity`
+  - A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/)
+- `podman`
+  - A generic configuration profile to be used with [Podman](https://podman.io/)
+- `shifter`
+  - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/)
+- `charliecloud`
+  - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/)
+- `apptainer`
+  - A generic configuration profile to be used with [Apptainer](https://apptainer.org/)
+- `conda`
+  - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter, Charliecloud, or Apptainer.
+
+### `-resume`
+
+Specify this when restarting a pipeline. Nextflow will use cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. For input to be considered the same, not only the names must be identical but the files' contents as well. For more info about this parameter, see [this blog post](https://www.nextflow.io/blog/2019/demystifying-nextflow-resume.html).
+
+You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names.
+
+### `-c`
+
+Specify the path to a specific config file (this is a core Nextflow command). See the [nf-core website documentation](https://nf-co.re/usage/configuration) for more information.
+
+## Custom configuration
+
+### Resource requests
+
+Whilst the default requirements set within the pipeline will hopefully work for most people and with most input data, you may find that you want to customise the compute resources that the pipeline requests. Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with any of the error codes specified [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L18) it will automatically be resubmitted with higher requests (2 x original, then 3 x original). If it still fails after the third attempt then the pipeline execution is stopped.
+
+To change the resource requests, please see the [max resources](https://nf-co.re/docs/usage/configuration#max-resources) and [tuning workflow resources](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources) section of the nf-core website.
+
+### Custom Containers
+
+In some cases you may wish to change which container or conda environment a step of the pipeline uses for a particular tool. By default nf-core pipelines use containers and software from the [biocontainers](https://biocontainers.pro/) or [bioconda](https://bioconda.github.io/) projects. However in some cases the pipeline specified version maybe out of date.
+
+To use a different container from the default container or conda environment specified in a pipeline, please see the [updating tool versions](https://nf-co.re/docs/usage/configuration#updating-tool-versions) section of the nf-core website.
+
+### Custom Tool Arguments
+
+A pipeline might not always support every possible argument or option of a particular tool used in pipeline. Fortunately, nf-core pipelines provide some freedom to users to insert additional parameters that the pipeline does not include by default.
+
+To learn how to provide additional arguments to a particular tool of the pipeline, please see the [customising tool arguments](https://nf-co.re/docs/usage/configuration#customising-tool-arguments) section of the nf-core website.
+
+### nf-core/configs
+
+In most cases, you will only need to create a custom config as a one-off but if you and others within your organisation are likely to be running nf-core pipelines regularly and need to use the same settings regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter. You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile.
+
+See the main [Nextflow documentation](https://www.nextflow.io/docs/latest/config.html) for more information about creating your own configuration files.
+
+If you have any questions or issues please send us a message on [Slack](https://nf-co.re/join/slack) on the [`#configs` channel](https://nfcore.slack.com/channels/configs).
+
+## Azure Resource Requests
+
+To be used with the `azurebatch` profile by specifying the `-profile azurebatch`.
+We recommend providing a compute `params.vm_type` of `Standard_D16_v3` VMs by default but these options can be changed if required.
+
+Note that the choice of VM size depends on your quota and the overall workload during the analysis.
+For a thorough list, please refer the [Azure Sizes for virtual machines in Azure](https://docs.microsoft.com/en-us/azure/virtual-machines/sizes).
+
+## Running in the background
+
+Nextflow handles job submissions and supervises the running jobs. The Nextflow process must run until the pipeline is finished.
+
+The Nextflow `-bg` flag launches Nextflow in the background, detached from your terminal so that the workflow does not stop if you log out of your session. The logs are saved to a file.
+
+Alternatively, you can use `screen` / `tmux` or similar tool to create a detached session which you can log back into at a later time.
+Some HPC setups also allow you to run nextflow within a cluster job submitted your job scheduler (from where it submits more jobs).
+
+## Nextflow memory requirements
+
+In some cases, the Nextflow Java virtual machines can start to request a large amount of memory.
+We recommend adding the following line to your environment to limit this (typically in `~/.bashrc` or `~./bash_profile`):
+
+```bash
+NXF_OPTS='-Xms1g -Xmx4g'
+```
diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
new file mode 100755
index 00000000..408951ae
--- /dev/null
+++ b/lib/NfcoreTemplate.groovy
@@ -0,0 +1,336 @@
+//
+// This file holds several functions used within the nf-core pipeline template.
+//
+
+import org.yaml.snakeyaml.Yaml
+
+class NfcoreTemplate {
+
+    //
+    // Check AWS Batch related parameters have been specified correctly
+    //
+    public static void awsBatch(workflow, params) {
+        if (workflow.profile.contains('awsbatch')) {
+            // Check params.awsqueue and params.awsregion have been set if running on AWSBatch
+            assert (params.awsqueue && params.awsregion) : "Specify correct --awsqueue and --awsregion parameters on AWSBatch!"
+            // Check outdir paths to be S3 buckets if running on AWSBatch
+            assert params.outdir.startsWith('s3:')       : "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!"
+        }
+    }
+
+    //
+    //  Warn if a -profile or Nextflow config has not been provided to run the pipeline
+    //
+    public static void checkConfigProvided(workflow, log) {
+        if (workflow.profile == 'standard' && workflow.configFiles.size() <= 1) {
+            log.warn "[$workflow.manifest.name] You are attempting to run the pipeline without any custom configuration!\n\n" +
+                    "This will be dependent on your local compute environment but can be achieved via one or more of the following:\n" +
+                    "   (1) Using an existing pipeline profile e.g. `-profile docker` or `-profile singularity`\n" +
+                    "   (2) Using an existing nf-core/configs for your Institution e.g. `-profile crick` or `-profile uppmax`\n" +
+                    "   (3) Using your own local custom config e.g. `-c /path/to/your/custom.config`\n\n" +
+                    "Please refer to the quick start section and usage docs for the pipeline.\n "
+        }
+    }
+
+    //
+    // Generate version string
+    //
+    public static String version(workflow) {
+        String version_string = ""
+
+        if (workflow.manifest.version) {
+            def prefix_v = workflow.manifest.version[0] != 'v' ? 'v' : ''
+            version_string += "${prefix_v}${workflow.manifest.version}"
+        }
+
+        if (workflow.commitId) {
+            def git_shortsha = workflow.commitId.substring(0, 7)
+            version_string += "-g${git_shortsha}"
+        }
+
+        return version_string
+    }
+
+    //
+    // Construct and send completion email
+    //
+    public static void email(workflow, params, summary_params, projectDir, log, multiqc_report=[]) {
+
+        // Set up the e-mail variables
+        def subject = "[$workflow.manifest.name] Successful: $workflow.runName"
+        if (!workflow.success) {
+            subject = "[$workflow.manifest.name] FAILED: $workflow.runName"
+        }
+
+        def summary = [:]
+        for (group in summary_params.keySet()) {
+            summary << summary_params[group]
+        }
+
+        def misc_fields = [:]
+        misc_fields['Date Started']              = workflow.start
+        misc_fields['Date Completed']            = workflow.complete
+        misc_fields['Pipeline script file path'] = workflow.scriptFile
+        misc_fields['Pipeline script hash ID']   = workflow.scriptId
+        if (workflow.repository) misc_fields['Pipeline repository Git URL']    = workflow.repository
+        if (workflow.commitId)   misc_fields['Pipeline repository Git Commit'] = workflow.commitId
+        if (workflow.revision)   misc_fields['Pipeline Git branch/tag']        = workflow.revision
+        misc_fields['Nextflow Version']           = workflow.nextflow.version
+        misc_fields['Nextflow Build']             = workflow.nextflow.build
+        misc_fields['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp
+
+        def email_fields = [:]
+        email_fields['version']      = NfcoreTemplate.version(workflow)
+        email_fields['runName']      = workflow.runName
+        email_fields['success']      = workflow.success
+        email_fields['dateComplete'] = workflow.complete
+        email_fields['duration']     = workflow.duration
+        email_fields['exitStatus']   = workflow.exitStatus
+        email_fields['errorMessage'] = (workflow.errorMessage ?: 'None')
+        email_fields['errorReport']  = (workflow.errorReport ?: 'None')
+        email_fields['commandLine']  = workflow.commandLine
+        email_fields['projectDir']   = workflow.projectDir
+        email_fields['summary']      = summary << misc_fields
+
+        // On success try attach the multiqc report
+        def mqc_report = null
+        try {
+            if (workflow.success) {
+                mqc_report = multiqc_report.getVal()
+                if (mqc_report.getClass() == ArrayList && mqc_report.size() >= 1) {
+                    if (mqc_report.size() > 1) {
+                        log.warn "[$workflow.manifest.name] Found multiple reports from process 'MULTIQC', will use only one"
+                    }
+                    mqc_report = mqc_report[0]
+                }
+            }
+        } catch (all) {
+            if (multiqc_report) {
+                log.warn "[$workflow.manifest.name] Could not attach MultiQC report to summary email"
+            }
+        }
+
+        // Check if we are only sending emails on failure
+        def email_address = params.email
+        if (!params.email && params.email_on_fail && !workflow.success) {
+            email_address = params.email_on_fail
+        }
+
+        // Render the TXT template
+        def engine       = new groovy.text.GStringTemplateEngine()
+        def tf           = new File("$projectDir/assets/email_template.txt")
+        def txt_template = engine.createTemplate(tf).make(email_fields)
+        def email_txt    = txt_template.toString()
+
+        // Render the HTML template
+        def hf            = new File("$projectDir/assets/email_template.html")
+        def html_template = engine.createTemplate(hf).make(email_fields)
+        def email_html    = html_template.toString()
+
+        // Render the sendmail template
+        def max_multiqc_email_size = (params.containsKey('max_multiqc_email_size') ? params.max_multiqc_email_size : 0) as nextflow.util.MemoryUnit
+        def smail_fields           = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ]
+        def sf                     = new File("$projectDir/assets/sendmail_template.txt")
+        def sendmail_template      = engine.createTemplate(sf).make(smail_fields)
+        def sendmail_html          = sendmail_template.toString()
+
+        // Send the HTML e-mail
+        Map colors = logColours(params.monochrome_logs)
+        if (email_address) {
+            try {
+                if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') }
+                // Try to send HTML e-mail using sendmail
+                [ 'sendmail', '-t' ].execute() << sendmail_html
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (sendmail)-"
+            } catch (all) {
+                // Catch failures and try with plaintext
+                def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ]
+                if ( mqc_report.size() <= max_multiqc_email_size.toBytes() ) {
+                    mail_cmd += [ '-A', mqc_report ]
+                }
+                mail_cmd.execute() << email_html
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (mail)-"
+            }
+        }
+
+        // Write summary e-mail HTML to a file
+        def output_d = new File("${params.outdir}/pipeline_info/")
+        if (!output_d.exists()) {
+            output_d.mkdirs()
+        }
+        def output_hf = new File(output_d, "pipeline_report.html")
+        output_hf.withWriter { w -> w << email_html }
+        def output_tf = new File(output_d, "pipeline_report.txt")
+        output_tf.withWriter { w -> w << email_txt }
+    }
+
+    //
+    // Construct and send a notification to a web server as JSON
+    // e.g. Microsoft Teams and Slack
+    //
+    public static void IM_notification(workflow, params, summary_params, projectDir, log) {
+        def hook_url = params.hook_url
+
+        def summary = [:]
+        for (group in summary_params.keySet()) {
+            summary << summary_params[group]
+        }
+
+        def misc_fields = [:]
+        misc_fields['start']                                = workflow.start
+        misc_fields['complete']                             = workflow.complete
+        misc_fields['scriptfile']                           = workflow.scriptFile
+        misc_fields['scriptid']                             = workflow.scriptId
+        if (workflow.repository) misc_fields['repository']  = workflow.repository
+        if (workflow.commitId)   misc_fields['commitid']    = workflow.commitId
+        if (workflow.revision)   misc_fields['revision']    = workflow.revision
+        misc_fields['nxf_version']                          = workflow.nextflow.version
+        misc_fields['nxf_build']                            = workflow.nextflow.build
+        misc_fields['nxf_timestamp']                        = workflow.nextflow.timestamp
+
+        def msg_fields = [:]
+        msg_fields['version']      = NfcoreTemplate.version(workflow)
+        msg_fields['runName']      = workflow.runName
+        msg_fields['success']      = workflow.success
+        msg_fields['dateComplete'] = workflow.complete
+        msg_fields['duration']     = workflow.duration
+        msg_fields['exitStatus']   = workflow.exitStatus
+        msg_fields['errorMessage'] = (workflow.errorMessage ?: 'None')
+        msg_fields['errorReport']  = (workflow.errorReport ?: 'None')
+        msg_fields['commandLine']  = workflow.commandLine.replaceFirst(/ +--hook_url +[^ ]+/, "")
+        msg_fields['projectDir']   = workflow.projectDir
+        msg_fields['summary']      = summary << misc_fields
+
+        // Render the JSON template
+        def engine       = new groovy.text.GStringTemplateEngine()
+        // Different JSON depending on the service provider
+        // Defaults to "Adaptive Cards" (https://adaptivecards.io), except Slack which has its own format
+        def json_path     = hook_url.contains("hooks.slack.com") ? "slackreport.json" : "adaptivecard.json"
+        def hf            = new File("$projectDir/assets/${json_path}")
+        def json_template = engine.createTemplate(hf).make(msg_fields)
+        def json_message  = json_template.toString()
+
+        // POST
+        def post = new URL(hook_url).openConnection();
+        post.setRequestMethod("POST")
+        post.setDoOutput(true)
+        post.setRequestProperty("Content-Type", "application/json")
+        post.getOutputStream().write(json_message.getBytes("UTF-8"));
+        def postRC = post.getResponseCode();
+        if (! postRC.equals(200)) {
+            log.warn(post.getErrorStream().getText());
+        }
+    }
+
+    //
+    // Print pipeline summary on completion
+    //
+    public static void summary(workflow, params, log) {
+        Map colors = logColours(params.monochrome_logs)
+        if (workflow.success) {
+            if (workflow.stats.ignoredCount == 0) {
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Pipeline completed successfully${colors.reset}-"
+            } else {
+                log.info "-${colors.purple}[$workflow.manifest.name]${colors.yellow} Pipeline completed successfully, but with errored process(es) ${colors.reset}-"
+            }
+        } else {
+            log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed with errors${colors.reset}-"
+        }
+    }
+
+    //
+    // ANSII Colours used for terminal logging
+    //
+    public static Map logColours(Boolean monochrome_logs) {
+        Map colorcodes = [:]
+
+        // Reset / Meta
+        colorcodes['reset']      = monochrome_logs ? '' : "\033[0m"
+        colorcodes['bold']       = monochrome_logs ? '' : "\033[1m"
+        colorcodes['dim']        = monochrome_logs ? '' : "\033[2m"
+        colorcodes['underlined'] = monochrome_logs ? '' : "\033[4m"
+        colorcodes['blink']      = monochrome_logs ? '' : "\033[5m"
+        colorcodes['reverse']    = monochrome_logs ? '' : "\033[7m"
+        colorcodes['hidden']     = monochrome_logs ? '' : "\033[8m"
+
+        // Regular Colors
+        colorcodes['black']      = monochrome_logs ? '' : "\033[0;30m"
+        colorcodes['red']        = monochrome_logs ? '' : "\033[0;31m"
+        colorcodes['green']      = monochrome_logs ? '' : "\033[0;32m"
+        colorcodes['yellow']     = monochrome_logs ? '' : "\033[0;33m"
+        colorcodes['blue']       = monochrome_logs ? '' : "\033[0;34m"
+        colorcodes['purple']     = monochrome_logs ? '' : "\033[0;35m"
+        colorcodes['cyan']       = monochrome_logs ? '' : "\033[0;36m"
+        colorcodes['white']      = monochrome_logs ? '' : "\033[0;37m"
+
+        // Bold
+        colorcodes['bblack']     = monochrome_logs ? '' : "\033[1;30m"
+        colorcodes['bred']       = monochrome_logs ? '' : "\033[1;31m"
+        colorcodes['bgreen']     = monochrome_logs ? '' : "\033[1;32m"
+        colorcodes['byellow']    = monochrome_logs ? '' : "\033[1;33m"
+        colorcodes['bblue']      = monochrome_logs ? '' : "\033[1;34m"
+        colorcodes['bpurple']    = monochrome_logs ? '' : "\033[1;35m"
+        colorcodes['bcyan']      = monochrome_logs ? '' : "\033[1;36m"
+        colorcodes['bwhite']     = monochrome_logs ? '' : "\033[1;37m"
+
+        // Underline
+        colorcodes['ublack']     = monochrome_logs ? '' : "\033[4;30m"
+        colorcodes['ured']       = monochrome_logs ? '' : "\033[4;31m"
+        colorcodes['ugreen']     = monochrome_logs ? '' : "\033[4;32m"
+        colorcodes['uyellow']    = monochrome_logs ? '' : "\033[4;33m"
+        colorcodes['ublue']      = monochrome_logs ? '' : "\033[4;34m"
+        colorcodes['upurple']    = monochrome_logs ? '' : "\033[4;35m"
+        colorcodes['ucyan']      = monochrome_logs ? '' : "\033[4;36m"
+        colorcodes['uwhite']     = monochrome_logs ? '' : "\033[4;37m"
+
+        // High Intensity
+        colorcodes['iblack']     = monochrome_logs ? '' : "\033[0;90m"
+        colorcodes['ired']       = monochrome_logs ? '' : "\033[0;91m"
+        colorcodes['igreen']     = monochrome_logs ? '' : "\033[0;92m"
+        colorcodes['iyellow']    = monochrome_logs ? '' : "\033[0;93m"
+        colorcodes['iblue']      = monochrome_logs ? '' : "\033[0;94m"
+        colorcodes['ipurple']    = monochrome_logs ? '' : "\033[0;95m"
+        colorcodes['icyan']      = monochrome_logs ? '' : "\033[0;96m"
+        colorcodes['iwhite']     = monochrome_logs ? '' : "\033[0;97m"
+
+        // Bold High Intensity
+        colorcodes['biblack']    = monochrome_logs ? '' : "\033[1;90m"
+        colorcodes['bired']      = monochrome_logs ? '' : "\033[1;91m"
+        colorcodes['bigreen']    = monochrome_logs ? '' : "\033[1;92m"
+        colorcodes['biyellow']   = monochrome_logs ? '' : "\033[1;93m"
+        colorcodes['biblue']     = monochrome_logs ? '' : "\033[1;94m"
+        colorcodes['bipurple']   = monochrome_logs ? '' : "\033[1;95m"
+        colorcodes['bicyan']     = monochrome_logs ? '' : "\033[1;96m"
+        colorcodes['biwhite']    = monochrome_logs ? '' : "\033[1;97m"
+
+        return colorcodes
+    }
+
+    //
+    // Does what is says on the tin
+    //
+    public static String dashedLine(monochrome_logs) {
+        Map colors = logColours(monochrome_logs)
+        return "-${colors.dim}----------------------------------------------------${colors.reset}-"
+    }
+
+    //
+    // nf-core logo
+    //
+    public static String logo(workflow, monochrome_logs) {
+        Map colors = logColours(monochrome_logs)
+        String workflow_version = NfcoreTemplate.version(workflow)
+        String.format(
+            """\n
+            ${dashedLine(monochrome_logs)}
+                                                    ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset}
+            ${colors.blue}        ___     __   __   __   ___     ${colors.green}/,-._.--~\'${colors.reset}
+            ${colors.blue}  |\\ | |__  __ /  ` /  \\ |__) |__         ${colors.yellow}}  {${colors.reset}
+            ${colors.blue}  | \\| |       \\__, \\__/ |  \\ |___     ${colors.green}\\`-._,-`-,${colors.reset}
+                                                    ${colors.green}`._,._,\'${colors.reset}
+            ${colors.purple}  ${workflow.manifest.name} ${workflow_version}${colors.reset}
+            ${dashedLine(monochrome_logs)}
+            """.stripIndent()
+        )
+    }
+}
diff --git a/lib/Utils.groovy b/lib/Utils.groovy
new file mode 100644
index 00000000..8d030f4e
--- /dev/null
+++ b/lib/Utils.groovy
@@ -0,0 +1,47 @@
+//
+// This file holds several Groovy functions that could be useful for any Nextflow pipeline
+//
+
+import org.yaml.snakeyaml.Yaml
+
+class Utils {
+
+    //
+    // When running with -profile conda, warn if channels have not been set-up appropriately
+    //
+    public static void checkCondaChannels(log) {
+        Yaml parser = new Yaml()
+        def channels = []
+        try {
+            def config = parser.load("conda config --show channels".execute().text)
+            channels = config.channels
+        } catch(NullPointerException | IOException e) {
+            log.warn "Could not verify conda channel configuration."
+            return
+        }
+
+        // Check that all channels are present
+        // This channel list is ordered by required channel priority.
+        def required_channels_in_order = ['conda-forge', 'bioconda', 'defaults']
+        def channels_missing = ((required_channels_in_order as Set) - (channels as Set)) as Boolean
+
+        // Check that they are in the right order
+        def channel_priority_violation = false
+        def n = required_channels_in_order.size()
+        for (int i = 0; i < n - 1; i++) {
+            channel_priority_violation |= !(channels.indexOf(required_channels_in_order[i]) < channels.indexOf(required_channels_in_order[i+1]))
+        }
+
+        if (channels_missing | channel_priority_violation) {
+            log.warn "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" +
+                "  There is a problem with your Conda configuration!\n\n" +
+                "  You will need to set-up the conda-forge and bioconda channels correctly.\n" +
+                "  Please refer to https://bioconda.github.io/\n" +
+                "  The observed channel order is \n" +
+                "  ${channels}\n" +
+                "  but the following channel order is required:\n" +
+                "  ${required_channels_in_order}\n" +
+                "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~"
+        }
+    }
+}
diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy
new file mode 100755
index 00000000..86f987ea
--- /dev/null
+++ b/lib/WorkflowMain.groovy
@@ -0,0 +1,63 @@
+//
+// This file holds several functions specific to the main.nf workflow in the nf-core/rnadeseq pipeline
+//
+
+import nextflow.Nextflow
+
+class WorkflowMain {
+
+    //
+    // Citation string for pipeline
+    //
+    public static String citation(workflow) {
+        return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" +
+            // TODO nf-core: Add Zenodo DOI for pipeline after first release
+            //"* The pipeline\n" +
+            //"  https://doi.org/10.5281/zenodo.XXXXXXX\n\n" +
+            "* The nf-core framework\n" +
+            "  https://doi.org/10.1038/s41587-020-0439-x\n\n" +
+            "* Software dependencies\n" +
+            "  https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md"
+    }
+
+
+    //
+    // Validate parameters and print summary to screen
+    //
+    public static void initialise(workflow, params, log) {
+
+        // Print workflow version and exit on --version
+        if (params.version) {
+            String workflow_version = NfcoreTemplate.version(workflow)
+            log.info "${workflow.manifest.name} ${workflow_version}"
+            System.exit(0)
+        }
+
+        // Check that a -profile or Nextflow config has been provided to run the pipeline
+        NfcoreTemplate.checkConfigProvided(workflow, log)
+
+        // Check that conda channels are set-up correctly
+        if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) {
+            Utils.checkCondaChannels(log)
+        }
+
+        // Check AWS batch settings
+        NfcoreTemplate.awsBatch(workflow, params)
+
+        // Check input has been provided
+        if (!params.input) {
+            Nextflow.error("Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'")
+        }
+    }
+    //
+    // Get attribute from genome config file e.g. fasta
+    //
+    public static Object getGenomeAttribute(params, attribute) {
+        if (params.genomes && params.genome && params.genomes.containsKey(params.genome)) {
+            if (params.genomes[ params.genome ].containsKey(attribute)) {
+                return params.genomes[ params.genome ][ attribute ]
+            }
+        }
+        return null
+    }
+}
diff --git a/lib/WorkflowRnadeseq.groovy b/lib/WorkflowRnadeseq.groovy
new file mode 100755
index 00000000..055f93c8
--- /dev/null
+++ b/lib/WorkflowRnadeseq.groovy
@@ -0,0 +1,122 @@
+//
+// This file holds several functions specific to the workflow/rnadeseq.nf in the nf-core/rnadeseq pipeline
+//
+
+import nextflow.Nextflow
+import groovy.text.SimpleTemplateEngine
+
+class WorkflowRnadeseq {
+
+    //
+    // Check and validate parameters
+    //
+    public static void initialise(params, log) {
+
+        genomeExistsError(params, log)
+
+
+        if (!params.fasta) {
+            Nextflow.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file."
+        }
+    }
+
+    //
+    // Get workflow summary for MultiQC
+    //
+    public static String paramsSummaryMultiqc(workflow, summary) {
+        String summary_section = ''
+        for (group in summary.keySet()) {
+            def group_params = summary.get(group)  // This gets the parameters of that particular group
+            if (group_params) {
+                summary_section += "    <p style=\"font-size:110%\"><b>$group</b></p>\n"
+                summary_section += "    <dl class=\"dl-horizontal\">\n"
+                for (param in group_params.keySet()) {
+                    summary_section += "        <dt>$param</dt><dd><samp>${group_params.get(param) ?: '<span style=\"color:#999999;\">N/A</a>'}</samp></dd>\n"
+                }
+                summary_section += "    </dl>\n"
+            }
+        }
+
+        String yaml_file_text  = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n"
+        yaml_file_text        += "description: ' - this information is collected when the pipeline is started.'\n"
+        yaml_file_text        += "section_name: '${workflow.manifest.name} Workflow Summary'\n"
+        yaml_file_text        += "section_href: 'https://github.com/${workflow.manifest.name}'\n"
+        yaml_file_text        += "plot_type: 'html'\n"
+        yaml_file_text        += "data: |\n"
+        yaml_file_text        += "${summary_section}"
+        return yaml_file_text
+    }
+
+    //
+    // Generate methods description for MultiQC
+    //
+
+    public static String toolCitationText(params) {
+
+        // TODO Optionally add in-text citation tools to this list.
+        // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "",
+        // Uncomment function in methodsDescriptionText to render in MultiQC report
+        def citation_text = [
+                "Tools used in the workflow included:",
+                "FastQC (Andrews 2010),",
+                "MultiQC (Ewels et al. 2016)",
+                "."
+            ].join(' ').trim()
+
+        return citation_text
+    }
+
+    public static String toolBibliographyText(params) {
+
+        // TODO Optionally add bibliographic entries to this list.
+        // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "<li>Author (2023) Pub name, Journal, DOI</li>" : "",
+        // Uncomment function in methodsDescriptionText to render in MultiQC report
+        def reference_text = [
+                "<li>Andrews S, (2010) FastQC, URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/).</li>",
+                "<li>Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: /10.1093/bioinformatics/btw354</li>"
+            ].join(' ').trim()
+
+        return reference_text
+    }
+
+    public static String methodsDescriptionText(run_workflow, mqc_methods_yaml, params) {
+        // Convert  to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file
+        def meta = [:]
+        meta.workflow = run_workflow.toMap()
+        meta["manifest_map"] = run_workflow.manifest.toMap()
+
+        // Pipeline DOI
+        meta["doi_text"] = meta.manifest_map.doi ? "(doi: <a href=\'https://doi.org/${meta.manifest_map.doi}\'>${meta.manifest_map.doi}</a>)" : ""
+        meta["nodoi_text"] = meta.manifest_map.doi ? "": "<li>If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used. </li>"
+
+        // Tool references
+        meta["tool_citations"] = ""
+        meta["tool_bibliography"] = ""
+
+        // TODO Only uncomment below if logic in toolCitationText/toolBibliographyText has been filled!
+        //meta["tool_citations"] = toolCitationText(params).replaceAll(", \\.", ".").replaceAll("\\. \\.", ".").replaceAll(", \\.", ".")
+        //meta["tool_bibliography"] = toolBibliographyText(params)
+
+
+        def methods_text = mqc_methods_yaml.text
+
+        def engine =  new SimpleTemplateEngine()
+        def description_html = engine.createTemplate(methods_text).make(meta)
+
+        return description_html
+    }
+
+    //
+    // Exit pipeline if incorrect --genome key provided
+    //
+    private static void genomeExistsError(params, log) {
+        if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) {
+            def error_string = "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" +
+                "  Genome '${params.genome}' not found in any config files provided to the pipeline.\n" +
+                "  Currently, the available genome keys are:\n" +
+                "  ${params.genomes.keySet().join(", ")}\n" +
+                "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~"
+            Nextflow.error(error_string)
+        }
+    }
+}
diff --git a/lib/nfcore_external_java_deps.jar b/lib/nfcore_external_java_deps.jar
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HcmV?d00001

diff --git a/main.nf b/main.nf
new file mode 100644
index 00000000..71d6b6c6
--- /dev/null
+++ b/main.nf
@@ -0,0 +1,79 @@
+#!/usr/bin/env nextflow
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Github : https://github.com/nf-core/rnadeseq
+    Website: https://nf-co.re/rnadeseq
+    Slack  : https://nfcore.slack.com/channels/rnadeseq
+----------------------------------------------------------------------------------------
+*/
+
+nextflow.enable.dsl = 2
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    GENOME PARAMETER VALUES
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+params.fasta = WorkflowMain.getGenomeAttribute(params, 'fasta')
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    VALIDATE & PRINT PARAMETER SUMMARY
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+include { validateParameters; paramsHelp } from 'plugin/nf-validation'
+
+// Print help message if needed
+if (params.help) {
+    def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs)
+    def citation = '\n' + WorkflowMain.citation(workflow) + '\n'
+    def String command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --genome GRCh37 -profile docker"
+    log.info logo + paramsHelp(command) + citation + NfcoreTemplate.dashedLine(params.monochrome_logs)
+    System.exit(0)
+}
+
+// Validate input parameters
+if (params.validate_params) {
+    validateParameters()
+}
+
+WorkflowMain.initialise(workflow, params, log)
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    NAMED WORKFLOW FOR PIPELINE
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+include { RNADESEQ } from './workflows/rnadeseq'
+
+//
+// WORKFLOW: Run main nf-core/rnadeseq analysis pipeline
+//
+workflow NFCORE_RNADESEQ {
+    RNADESEQ ()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    RUN ALL WORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// WORKFLOW: Execute a single named workflow for the pipeline
+// See: https://github.com/nf-core/rnaseq/issues/619
+//
+workflow {
+    NFCORE_RNADESEQ ()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    THE END
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
diff --git a/modules.json b/modules.json
new file mode 100644
index 00000000..b4265295
--- /dev/null
+++ b/modules.json
@@ -0,0 +1,27 @@
+{
+    "name": "nf-core/rnadeseq",
+    "homePage": "https://github.com/nf-core/rnadeseq",
+    "repos": {
+        "https://github.com/nf-core/modules.git": {
+            "modules": {
+                "nf-core": {
+                    "custom/dumpsoftwareversions": {
+                        "branch": "master",
+                        "git_sha": "76cc4938c1f6ea5c7d83fed1eeffc146787f9543",
+                        "installed_by": ["modules"]
+                    },
+                    "fastqc": {
+                        "branch": "master",
+                        "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c",
+                        "installed_by": ["modules"]
+                    },
+                    "multiqc": {
+                        "branch": "master",
+                        "git_sha": "f2d63bd5b68925f98f572eed70993d205cc694b7",
+                        "installed_by": ["modules"]
+                    }
+                }
+            }
+        }
+    }
+}
diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf
new file mode 100644
index 00000000..05aa5263
--- /dev/null
+++ b/modules/local/samplesheet_check.nf
@@ -0,0 +1,31 @@
+process SAMPLESHEET_CHECK {
+    tag "$samplesheet"
+    label 'process_single'
+
+    conda "conda-forge::python=3.8.3"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/python:3.8.3' :
+        'biocontainers/python:3.8.3' }"
+
+    input:
+    path samplesheet
+
+    output:
+    path '*.csv'       , emit: csv
+    path "versions.yml", emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script: // This script is bundled with the pipeline, in nf-core/rnadeseq/bin/
+    """
+    check_samplesheet.py \\
+        $samplesheet \\
+        samplesheet.valid.csv
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        python: \$(python --version | sed 's/Python //g')
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/custom/dumpsoftwareversions/main.nf b/modules/nf-core/custom/dumpsoftwareversions/main.nf
new file mode 100644
index 00000000..800a6099
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/main.nf
@@ -0,0 +1,24 @@
+process CUSTOM_DUMPSOFTWAREVERSIONS {
+    label 'process_single'
+
+    // Requires `pyyaml` which does not have a dedicated container but is in the MultiQC container
+    conda "bioconda::multiqc=1.14"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/multiqc:1.14--pyhdfd78af_0' :
+        'quay.io/biocontainers/multiqc:1.14--pyhdfd78af_0' }"
+
+    input:
+    path versions
+
+    output:
+    path "software_versions.yml"    , emit: yml
+    path "software_versions_mqc.yml", emit: mqc_yml
+    path "versions.yml"             , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    template 'dumpsoftwareversions.py'
+}
diff --git a/modules/nf-core/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/custom/dumpsoftwareversions/meta.yml
new file mode 100644
index 00000000..c32657de
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/meta.yml
@@ -0,0 +1,36 @@
+# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/yaml-schema.json
+name: custom_dumpsoftwareversions
+description: Custom module used to dump software versions within the nf-core pipeline template
+keywords:
+  - custom
+  - dump
+  - version
+tools:
+  - custom:
+      description: Custom module used to dump software versions within the nf-core pipeline template
+      homepage: https://github.com/nf-core/tools
+      documentation: https://github.com/nf-core/tools
+      licence: ["MIT"]
+input:
+  - versions:
+      type: file
+      description: YML file containing software versions
+      pattern: "*.yml"
+
+output:
+  - yml:
+      type: file
+      description: Standard YML file containing software versions
+      pattern: "software_versions.yml"
+  - mqc_yml:
+      type: file
+      description: MultiQC custom content YML file containing software versions
+      pattern: "software_versions_mqc.yml"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+
+authors:
+  - "@drpatelh"
+  - "@grst"
diff --git a/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
new file mode 100755
index 00000000..e55b8d43
--- /dev/null
+++ b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py
@@ -0,0 +1,102 @@
+#!/usr/bin/env python
+
+
+"""Provide functions to merge multiple versions.yml files."""
+
+
+import platform
+from textwrap import dedent
+
+import yaml
+
+
+def _make_versions_html(versions):
+    """Generate a tabular HTML output of all versions for MultiQC."""
+    html = [
+        dedent(
+            """\\
+            <style>
+            #nf-core-versions tbody:nth-child(even) {
+                background-color: #f2f2f2;
+            }
+            </style>
+            <table class="table" style="width:100%" id="nf-core-versions">
+                <thead>
+                    <tr>
+                        <th> Process Name </th>
+                        <th> Software </th>
+                        <th> Version  </th>
+                    </tr>
+                </thead>
+            """
+        )
+    ]
+    for process, tmp_versions in sorted(versions.items()):
+        html.append("<tbody>")
+        for i, (tool, version) in enumerate(sorted(tmp_versions.items())):
+            html.append(
+                dedent(
+                    f"""\\
+                    <tr>
+                        <td><samp>{process if (i == 0) else ''}</samp></td>
+                        <td><samp>{tool}</samp></td>
+                        <td><samp>{version}</samp></td>
+                    </tr>
+                    """
+                )
+            )
+        html.append("</tbody>")
+    html.append("</table>")
+    return "\\n".join(html)
+
+
+def main():
+    """Load all version files and generate merged output."""
+    versions_this_module = {}
+    versions_this_module["${task.process}"] = {
+        "python": platform.python_version(),
+        "yaml": yaml.__version__,
+    }
+
+    with open("$versions") as f:
+        versions_by_process = yaml.load(f, Loader=yaml.BaseLoader) | versions_this_module
+
+    # aggregate versions by the module name (derived from fully-qualified process name)
+    versions_by_module = {}
+    for process, process_versions in versions_by_process.items():
+        module = process.split(":")[-1]
+        try:
+            if versions_by_module[module] != process_versions:
+                raise AssertionError(
+                    "We assume that software versions are the same between all modules. "
+                    "If you see this error-message it means you discovered an edge-case "
+                    "and should open an issue in nf-core/tools. "
+                )
+        except KeyError:
+            versions_by_module[module] = process_versions
+
+    versions_by_module["Workflow"] = {
+        "Nextflow": "$workflow.nextflow.version",
+        "$workflow.manifest.name": "$workflow.manifest.version",
+    }
+
+    versions_mqc = {
+        "id": "software_versions",
+        "section_name": "${workflow.manifest.name} Software Versions",
+        "section_href": "https://github.com/${workflow.manifest.name}",
+        "plot_type": "html",
+        "description": "are collected at run time from the software output.",
+        "data": _make_versions_html(versions_by_module),
+    }
+
+    with open("software_versions.yml", "w") as f:
+        yaml.dump(versions_by_module, f, default_flow_style=False)
+    with open("software_versions_mqc.yml", "w") as f:
+        yaml.dump(versions_mqc, f, default_flow_style=False)
+
+    with open("versions.yml", "w") as f:
+        yaml.dump(versions_this_module, f, default_flow_style=False)
+
+
+if __name__ == "__main__":
+    main()
diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf
new file mode 100644
index 00000000..9ae58381
--- /dev/null
+++ b/modules/nf-core/fastqc/main.nf
@@ -0,0 +1,51 @@
+process FASTQC {
+    tag "$meta.id"
+    label 'process_medium'
+
+    conda "bioconda::fastqc=0.11.9"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0' :
+        'quay.io/biocontainers/fastqc:0.11.9--0' }"
+
+    input:
+    tuple val(meta), path(reads)
+
+    output:
+    tuple val(meta), path("*.html"), emit: html
+    tuple val(meta), path("*.zip") , emit: zip
+    path  "versions.yml"           , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    def prefix = task.ext.prefix ?: "${meta.id}"
+    // Make list of old name and new name pairs to use for renaming in the bash while loop
+    def old_new_pairs = reads instanceof Path || reads.size() == 1 ? [[ reads, "${prefix}.${reads.extension}" ]] : reads.withIndex().collect { entry, index -> [ entry, "${prefix}_${index + 1}.${entry.extension}" ] }
+    def rename_to = old_new_pairs*.join(' ').join(' ')
+    def renamed_files = old_new_pairs.collect{ old_name, new_name -> new_name }.join(' ')
+    """
+    printf "%s %s\\n" $rename_to | while read old_name new_name; do
+        [ -f "\${new_name}" ] || ln -s \$old_name \$new_name
+    done
+    fastqc $args --threads $task.cpus $renamed_files
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
+    END_VERSIONS
+    """
+
+    stub:
+    def prefix = task.ext.prefix ?: "${meta.id}"
+    """
+    touch ${prefix}.html
+    touch ${prefix}.zip
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" )
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/fastqc/meta.yml b/modules/nf-core/fastqc/meta.yml
new file mode 100644
index 00000000..4da5bb5a
--- /dev/null
+++ b/modules/nf-core/fastqc/meta.yml
@@ -0,0 +1,52 @@
+name: fastqc
+description: Run FastQC on sequenced reads
+keywords:
+  - quality control
+  - qc
+  - adapters
+  - fastq
+tools:
+  - fastqc:
+      description: |
+        FastQC gives general quality metrics about your reads.
+        It provides information about the quality score distribution
+        across your reads, the per base sequence content (%A/C/G/T).
+        You get information about adapter contamination and other
+        overrepresented sequences.
+      homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/
+      documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/
+      licence: ["GPL-2.0-only"]
+input:
+  - meta:
+      type: map
+      description: |
+        Groovy Map containing sample information
+        e.g. [ id:'test', single_end:false ]
+  - reads:
+      type: file
+      description: |
+        List of input FastQ files of size 1 and 2 for single-end and paired-end data,
+        respectively.
+output:
+  - meta:
+      type: map
+      description: |
+        Groovy Map containing sample information
+        e.g. [ id:'test', single_end:false ]
+  - html:
+      type: file
+      description: FastQC report
+      pattern: "*_{fastqc.html}"
+  - zip:
+      type: file
+      description: FastQC report archive
+      pattern: "*_{fastqc.zip}"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+authors:
+  - "@drpatelh"
+  - "@grst"
+  - "@ewels"
+  - "@FelixKrueger"
diff --git a/modules/nf-core/multiqc/main.nf b/modules/nf-core/multiqc/main.nf
new file mode 100644
index 00000000..4b604749
--- /dev/null
+++ b/modules/nf-core/multiqc/main.nf
@@ -0,0 +1,53 @@
+process MULTIQC {
+    label 'process_single'
+
+    conda "bioconda::multiqc=1.14"
+    container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ?
+        'https://depot.galaxyproject.org/singularity/multiqc:1.14--pyhdfd78af_0' :
+        'quay.io/biocontainers/multiqc:1.14--pyhdfd78af_0' }"
+
+    input:
+    path  multiqc_files, stageAs: "?/*"
+    path(multiqc_config)
+    path(extra_multiqc_config)
+    path(multiqc_logo)
+
+    output:
+    path "*multiqc_report.html", emit: report
+    path "*_data"              , emit: data
+    path "*_plots"             , optional:true, emit: plots
+    path "versions.yml"        , emit: versions
+
+    when:
+    task.ext.when == null || task.ext.when
+
+    script:
+    def args = task.ext.args ?: ''
+    def config = multiqc_config ? "--config $multiqc_config" : ''
+    def extra_config = extra_multiqc_config ? "--config $extra_multiqc_config" : ''
+    """
+    multiqc \\
+        --force \\
+        $args \\
+        $config \\
+        $extra_config \\
+        .
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
+    END_VERSIONS
+    """
+
+    stub:
+    """
+    touch multiqc_data
+    touch multiqc_plots
+    touch multiqc_report.html
+
+    cat <<-END_VERSIONS > versions.yml
+    "${task.process}":
+        multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" )
+    END_VERSIONS
+    """
+}
diff --git a/modules/nf-core/multiqc/meta.yml b/modules/nf-core/multiqc/meta.yml
new file mode 100644
index 00000000..f93b5ee5
--- /dev/null
+++ b/modules/nf-core/multiqc/meta.yml
@@ -0,0 +1,56 @@
+# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/yaml-schema.json
+name: MultiQC
+description: Aggregate results from bioinformatics analyses across many samples into a single report
+keywords:
+  - QC
+  - bioinformatics tools
+  - Beautiful stand-alone HTML report
+tools:
+  - multiqc:
+      description: |
+        MultiQC searches a given directory for analysis logs and compiles a HTML report.
+        It's a general use tool, perfect for summarising the output from numerous bioinformatics tools.
+      homepage: https://multiqc.info/
+      documentation: https://multiqc.info/docs/
+      licence: ["GPL-3.0-or-later"]
+
+input:
+  - multiqc_files:
+      type: file
+      description: |
+        List of reports / files recognised by MultiQC, for example the html and zip output of FastQC
+  - multiqc_config:
+      type: file
+      description: Optional config yml for MultiQC
+      pattern: "*.{yml,yaml}"
+  - extra_multiqc_config:
+      type: file
+      description: Second optional config yml for MultiQC. Will override common sections in multiqc_config.
+      pattern: "*.{yml,yaml}"
+  - multiqc_logo:
+      type: file
+      description: Optional logo file for MultiQC
+      pattern: "*.{png}"
+
+output:
+  - report:
+      type: file
+      description: MultiQC report file
+      pattern: "multiqc_report.html"
+  - data:
+      type: directory
+      description: MultiQC data dir
+      pattern: "multiqc_data"
+  - plots:
+      type: file
+      description: Plots created by MultiQC
+      pattern: "*_data"
+  - versions:
+      type: file
+      description: File containing software versions
+      pattern: "versions.yml"
+authors:
+  - "@abhi18av"
+  - "@bunop"
+  - "@drpatelh"
+  - "@jfy133"
diff --git a/nextflow.config b/nextflow.config
new file mode 100644
index 00000000..a25c5b73
--- /dev/null
+++ b/nextflow.config
@@ -0,0 +1,269 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    nf-core/rnadeseq Nextflow config file
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    Default config options for all compute environments
+----------------------------------------------------------------------------------------
+*/
+
+// Global default params, used in configs
+params {
+
+    // TODO nf-core: Specify your pipeline's command line flags
+    // Input options
+    input                      = null
+    // References
+    genome                     = null
+    igenomes_base              = 's3://ngi-igenomes/igenomes'
+    igenomes_ignore            = false
+    
+
+    // MultiQC options
+    multiqc_config             = null
+    multiqc_title              = null
+    multiqc_logo               = null
+    max_multiqc_email_size     = '25.MB'
+    multiqc_methods_description = null
+
+    // Boilerplate options
+    outdir                     = null
+    publish_dir_mode           = 'copy'
+    email                      = null
+    email_on_fail              = null
+    plaintext_email            = false
+    monochrome_logs            = false
+    hook_url                   = null
+    help                       = false
+    version                    = false
+
+    // Config options
+    config_profile_name        = null
+    config_profile_description = null
+    custom_config_version      = 'master'
+    custom_config_base         = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}"
+    config_profile_contact     = null
+    config_profile_url         = null
+    
+
+    // Max resource options
+    // Defaults only, expecting to be overwritten
+    max_memory                 = '128.GB'
+    max_cpus                   = 16
+    max_time                   = '240.h'
+
+    // Schema validation default options
+    validationFailUnrecognisedParams = false
+    validationLenientMode            = false
+    validationSchemaIgnoreParams     = 'genomes'
+    validationShowHiddenParams       = false
+    validate_params                  = true
+
+}
+
+// Load base.config by default for all pipelines
+includeConfig 'conf/base.config'
+
+// Load nf-core custom profiles from different Institutions
+try {
+    includeConfig "${params.custom_config_base}/nfcore_custom.config"
+} catch (Exception e) {
+    System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config")
+}
+
+// Load nf-core/rnadeseq custom profiles from different institutions.
+// Warning: Uncomment only if a pipeline-specific instititutional config already exists on nf-core/configs!
+// try {
+//   includeConfig "${params.custom_config_base}/pipeline/rnadeseq.config"
+// } catch (Exception e) {
+//   System.err.println("WARNING: Could not load nf-core/config/rnadeseq profiles: ${params.custom_config_base}/pipeline/rnadeseq.config")
+// }
+profiles {
+    debug {
+        dumpHashes             = true
+        process.beforeScript   = 'echo $HOSTNAME'
+        cleanup                = false
+    }
+    conda {
+        conda.enabled          = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    mamba {
+        conda.enabled          = true
+        conda.useMamba         = true
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    docker {
+        docker.enabled         = true
+        docker.userEmulation   = true
+        conda.enabled          = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    arm {
+        docker.runOptions = '-u $(id -u):$(id -g) --platform=linux/amd64'
+    }
+    singularity {
+        singularity.enabled    = true
+        singularity.autoMounts = true
+        conda.enabled          = false
+        docker.enabled         = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    podman {
+        podman.enabled         = true
+        conda.enabled          = false
+        docker.enabled         = false
+        singularity.enabled    = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    shifter {
+        shifter.enabled        = true
+        conda.enabled          = false
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        charliecloud.enabled   = false
+        apptainer.enabled      = false
+    }
+    charliecloud {
+        charliecloud.enabled   = true
+        conda.enabled          = false
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        apptainer.enabled      = false
+    }
+    apptainer {
+        apptainer.enabled      = true
+        conda.enabled          = false
+        docker.enabled         = false
+        singularity.enabled    = false
+        podman.enabled         = false
+        shifter.enabled        = false
+        charliecloud.enabled   = false
+    }
+    gitpod {
+        executor.name          = 'local'
+        executor.cpus          = 16
+        executor.memory        = 60.GB
+    }
+    test      { includeConfig 'conf/test.config'      }
+    test_full { includeConfig 'conf/test_full.config' }
+}
+
+// Set default registry for Apptainer, Docker, Podman and Singularity independent of -profile
+// Will not be used unless Apptainer / Docker / Podman / Singularity are enabled
+// Set to your registry if you have a mirror of containers
+apptainer.registry   = 'quay.io'
+docker.registry      = 'quay.io'
+podman.registry      = 'quay.io'
+singularity.registry = 'quay.io'
+
+// Nextflow plugins
+plugins {
+    id 'nf-validation' // Validation of pipeline parameters and creation of an input channel from a sample sheet
+}
+
+// Load igenomes.config if required
+if (!params.igenomes_ignore) {
+    includeConfig 'conf/igenomes.config'
+} else {
+    params.genomes = [:]
+}
+// Export these variables to prevent local Python/R libraries from conflicting with those in the container
+// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container.
+// See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable.
+
+env {
+    PYTHONNOUSERSITE = 1
+    R_PROFILE_USER   = "/.Rprofile"
+    R_ENVIRON_USER   = "/.Renviron"
+    JULIA_DEPOT_PATH = "/usr/local/share/julia"
+}
+
+// Capture exit codes from upstream processes when piping
+process.shell = ['/bin/bash', '-euo', 'pipefail']
+
+def trace_timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss')
+timeline {
+    enabled = true
+    file    = "${params.outdir}/pipeline_info/execution_timeline_${trace_timestamp}.html"
+}
+report {
+    enabled = true
+    file    = "${params.outdir}/pipeline_info/execution_report_${trace_timestamp}.html"
+}
+trace {
+    enabled = true
+    file    = "${params.outdir}/pipeline_info/execution_trace_${trace_timestamp}.txt"
+}
+dag {
+    enabled = true
+    file    = "${params.outdir}/pipeline_info/pipeline_dag_${trace_timestamp}.html"
+}
+
+manifest {
+    name            = 'nf-core/rnadeseq'
+    author          = """Gisela Gabernet, Silvia Morini, Oskar Wacker, QBiC"""
+    homePage        = 'https://github.com/nf-core/rnadeseq'
+    description     = """Differential gene expression analysis and pathway analysis of RNAseq data"""
+    mainScript      = 'main.nf'
+    nextflowVersion = '!>=23.04.0'
+    version         = '2.2dev'
+    doi             = ''
+}
+
+// Load modules.config for DSL2 module specific options
+includeConfig 'conf/modules.config'
+
+// Function to ensure that resource requirements don't go beyond
+// a maximum limit
+def check_max(obj, type) {
+    if (type == 'memory') {
+        try {
+            if (obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1)
+                return params.max_memory as nextflow.util.MemoryUnit
+            else
+                return obj
+        } catch (all) {
+            println "   ### ERROR ###   Max memory '${params.max_memory}' is not valid! Using default value: $obj"
+            return obj
+        }
+    } else if (type == 'time') {
+        try {
+            if (obj.compareTo(params.max_time as nextflow.util.Duration) == 1)
+                return params.max_time as nextflow.util.Duration
+            else
+                return obj
+        } catch (all) {
+            println "   ### ERROR ###   Max time '${params.max_time}' is not valid! Using default value: $obj"
+            return obj
+        }
+    } else if (type == 'cpus') {
+        try {
+            return Math.min( obj, params.max_cpus as int )
+        } catch (all) {
+            println "   ### ERROR ###   Max cpus '${params.max_cpus}' is not valid! Using default value: $obj"
+            return obj
+        }
+    }
+}
diff --git a/nextflow_schema.json b/nextflow_schema.json
new file mode 100644
index 00000000..c459e4b0
--- /dev/null
+++ b/nextflow_schema.json
@@ -0,0 +1,303 @@
+{
+    "$schema": "http://json-schema.org/draft-07/schema",
+    "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/nextflow_schema.json",
+    "title": "nf-core/rnadeseq pipeline parameters",
+    "description": "Differential gene expression analysis and pathway analysis of RNAseq data",
+    "type": "object",
+    "definitions": {
+        "input_output_options": {
+            "title": "Input/output options",
+            "type": "object",
+            "fa_icon": "fas fa-terminal",
+            "description": "Define where the pipeline should find input data and save output data.",
+            "required": ["input", "outdir"],
+            "properties": {
+                "input": {
+                    "type": "string",
+                    "format": "file-path",
+                    "exists": true,
+                    "mimetype": "text/csv",
+                    "pattern": "^\\S+\\.csv$",
+                    "description": "Path to comma-separated file containing information about the samples in the experiment.",
+                    "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row. See [usage docs](https://nf-co.re/rnadeseq/usage#samplesheet-input).",
+                    "fa_icon": "fas fa-file-csv"
+                },
+                "outdir": {
+                    "type": "string",
+                    "format": "directory-path",
+                    "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.",
+                    "fa_icon": "fas fa-folder-open"
+                },
+                "email": {
+                    "type": "string",
+                    "description": "Email address for completion summary.",
+                    "fa_icon": "fas fa-envelope",
+                    "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.",
+                    "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$"
+                },
+                "multiqc_title": {
+                    "type": "string",
+                    "description": "MultiQC report title. Printed as page header, used for filename if not otherwise specified.",
+                    "fa_icon": "fas fa-file-signature"
+                }
+            }
+        },
+        "reference_genome_options": {
+            "title": "Reference genome options",
+            "type": "object",
+            "fa_icon": "fas fa-dna",
+            "description": "Reference genome related files and options required for the workflow.",
+            "properties": {
+                "genome": {
+                    "type": "string",
+                    "description": "Name of iGenomes reference.",
+                    "fa_icon": "fas fa-book",
+                    "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files e.g. `--genome GRCh38`. \n\nSee the [nf-core website docs](https://nf-co.re/usage/reference_genomes) for more details."
+                },
+                "fasta": {
+                    "type": "string",
+                    "format": "file-path",
+                    "exists": true,
+                    "mimetype": "text/plain",
+                    "pattern": "^\\S+\\.fn?a(sta)?(\\.gz)?$",
+                    "description": "Path to FASTA genome file.",
+                    "help_text": "This parameter is *mandatory* if `--genome` is not specified. If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs.",
+                    "fa_icon": "far fa-file-code"
+                },
+                "igenomes_base": {
+                    "type": "string",
+                    "format": "directory-path",
+                    "description": "Directory / URL base for iGenomes references.",
+                    "default": "s3://ngi-igenomes/igenomes",
+                    "fa_icon": "fas fa-cloud-download-alt",
+                    "hidden": true
+                },
+                "igenomes_ignore": {
+                    "type": "boolean",
+                    "description": "Do not load the iGenomes reference config.",
+                    "fa_icon": "fas fa-ban",
+                    "hidden": true,
+                    "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`."
+                }
+            }
+        },
+        "institutional_config_options": {
+            "title": "Institutional config options",
+            "type": "object",
+            "fa_icon": "fas fa-university",
+            "description": "Parameters used to describe centralised config profiles. These should not be edited.",
+            "help_text": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.",
+            "properties": {
+                "custom_config_version": {
+                    "type": "string",
+                    "description": "Git commit id for Institutional configs.",
+                    "default": "master",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "custom_config_base": {
+                    "type": "string",
+                    "description": "Base directory for Institutional configs.",
+                    "default": "https://raw.githubusercontent.com/nf-core/configs/master",
+                    "hidden": true,
+                    "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.",
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_name": {
+                    "type": "string",
+                    "description": "Institutional config name.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_description": {
+                    "type": "string",
+                    "description": "Institutional config description.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_contact": {
+                    "type": "string",
+                    "description": "Institutional config contact information.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                },
+                "config_profile_url": {
+                    "type": "string",
+                    "description": "Institutional config URL link.",
+                    "hidden": true,
+                    "fa_icon": "fas fa-users-cog"
+                }
+            }
+        },
+        "max_job_request_options": {
+            "title": "Max job request options",
+            "type": "object",
+            "fa_icon": "fab fa-acquisitions-incorporated",
+            "description": "Set the top limit for requested resources for any single job.",
+            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
+            "properties": {
+                "max_cpus": {
+                    "type": "integer",
+                    "description": "Maximum number of CPUs that can be requested for any single job.",
+                    "default": 16,
+                    "fa_icon": "fas fa-microchip",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`"
+                },
+                "max_memory": {
+                    "type": "string",
+                    "description": "Maximum amount of memory that can be requested for any single job.",
+                    "default": "128.GB",
+                    "fa_icon": "fas fa-memory",
+                    "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`"
+                },
+                "max_time": {
+                    "type": "string",
+                    "description": "Maximum amount of time that can be requested for any single job.",
+                    "default": "240.h",
+                    "fa_icon": "far fa-clock",
+                    "pattern": "^(\\d+\\.?\\s*(s|m|h|d|day)\\s*)+$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`"
+                }
+            }
+        },
+        "generic_options": {
+            "title": "Generic options",
+            "type": "object",
+            "fa_icon": "fas fa-file-import",
+            "description": "Less common options for the pipeline, typically set in a config file.",
+            "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.",
+            "properties": {
+                "help": {
+                    "type": "boolean",
+                    "description": "Display help text.",
+                    "fa_icon": "fas fa-question-circle",
+                    "default": false,
+                    "hidden": true
+                },
+                "version": {
+                    "type": "boolean",
+                    "description": "Display version and exit.",
+                    "fa_icon": "fas fa-question-circle",
+                    "default": false,
+                    "hidden": true
+                },
+                "publish_dir_mode": {
+                    "type": "string",
+                    "default": "copy",
+                    "description": "Method used to save pipeline results to output directory.",
+                    "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.",
+                    "fa_icon": "fas fa-copy",
+                    "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"],
+                    "hidden": true
+                },
+                "email_on_fail": {
+                    "type": "string",
+                    "description": "Email address for completion summary, only when pipeline fails.",
+                    "fa_icon": "fas fa-exclamation-triangle",
+                    "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$",
+                    "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.",
+                    "hidden": true
+                },
+                "plaintext_email": {
+                    "type": "boolean",
+                    "description": "Send plain-text email instead of HTML.",
+                    "fa_icon": "fas fa-remove-format",
+                    "default": false,
+                    "hidden": true
+                },
+                "max_multiqc_email_size": {
+                    "type": "string",
+                    "description": "File size limit when attaching MultiQC reports to summary emails.",
+                    "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$",
+                    "default": "25.MB",
+                    "fa_icon": "fas fa-file-upload",
+                    "hidden": true
+                },
+                "monochrome_logs": {
+                    "type": "boolean",
+                    "description": "Do not use coloured log outputs.",
+                    "fa_icon": "fas fa-palette",
+                    "default": false,
+                    "hidden": true
+                },
+                "hook_url": {
+                    "type": "string",
+                    "description": "Incoming hook URL for messaging service",
+                    "fa_icon": "fas fa-people-group",
+                    "help_text": "Incoming hook URL for messaging service. Currently, MS Teams and Slack are supported.",
+                    "hidden": true
+                },
+                "multiqc_config": {
+                    "type": "string",
+                    "format": "file-path",
+                    "description": "Custom config file to supply to MultiQC.",
+                    "fa_icon": "fas fa-cog",
+                    "hidden": true
+                },
+                "multiqc_logo": {
+                    "type": "string",
+                    "description": "Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file",
+                    "fa_icon": "fas fa-image",
+                    "hidden": true
+                },
+                "multiqc_methods_description": {
+                    "type": "string",
+                    "description": "Custom MultiQC yaml file containing HTML including a methods description.",
+                    "fa_icon": "fas fa-cog"
+                },
+                "validate_params": {
+                    "type": "boolean",
+                    "description": "Boolean whether to validate parameters against the schema at runtime",
+                    "default": true,
+                    "fa_icon": "fas fa-check-square",
+                    "hidden": true
+                },
+                "validationShowHiddenParams": {
+                    "type": "boolean",
+                    "fa_icon": "far fa-eye-slash",
+                    "description": "Show all params when using `--help`",
+                    "default": false,
+                    "hidden": true,
+                    "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters."
+                },
+                "validationFailUnrecognisedParams": {
+                    "type": "boolean",
+                    "fa_icon": "far fa-check-circle",
+                    "description": "Validation of parameters fails when an unrecognised parameter is found.",
+                    "default": false,
+                    "hidden": true,
+                    "help_text": "By default, when an unrecognised parameter is found, it returns a warinig."
+                },
+                "validationLenientMode": {
+                    "type": "boolean",
+                    "fa_icon": "far fa-check-circle",
+                    "description": "Validation of parameters in lenient more.",
+                    "default": false,
+                    "hidden": true,
+                    "help_text": "Allows string values that are parseable as numbers or booleans. For further information see [JSONSchema docs](https://github.com/everit-org/json-schema#lenient-mode)."
+                }
+            }
+        }
+    },
+    "allOf": [
+        {
+            "$ref": "#/definitions/input_output_options"
+        },
+        {
+            "$ref": "#/definitions/reference_genome_options"
+        },
+        {
+            "$ref": "#/definitions/institutional_config_options"
+        },
+        {
+            "$ref": "#/definitions/max_job_request_options"
+        },
+        {
+            "$ref": "#/definitions/generic_options"
+        }
+    ]
+}
diff --git a/pyproject.toml b/pyproject.toml
new file mode 100644
index 00000000..0d62beb6
--- /dev/null
+++ b/pyproject.toml
@@ -0,0 +1,10 @@
+# Config file for Python. Mostly used to configure linting of bin/check_samplesheet.py with Black.
+# Should be kept the same as nf-core/tools to avoid fighting with template synchronisation.
+[tool.black]
+line-length = 120
+target_version = ["py37", "py38", "py39", "py310"]
+
+[tool.isort]
+profile = "black"
+known_first_party = ["nf_core"]
+multi_line_output = 3
diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf
new file mode 100644
index 00000000..0aecf87f
--- /dev/null
+++ b/subworkflows/local/input_check.nf
@@ -0,0 +1,44 @@
+//
+// Check input samplesheet and get read channels
+//
+
+include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check'
+
+workflow INPUT_CHECK {
+    take:
+    samplesheet // file: /path/to/samplesheet.csv
+
+    main:
+    SAMPLESHEET_CHECK ( samplesheet )
+        .csv
+        .splitCsv ( header:true, sep:',' )
+        .map { create_fastq_channel(it) }
+        .set { reads }
+
+    emit:
+    reads                                     // channel: [ val(meta), [ reads ] ]
+    versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ]
+}
+
+// Function to get list of [ meta, [ fastq_1, fastq_2 ] ]
+def create_fastq_channel(LinkedHashMap row) {
+    // create meta map
+    def meta = [:]
+    meta.id         = row.sample
+    meta.single_end = row.single_end.toBoolean()
+
+    // add path(s) of the fastq file(s) to the meta map
+    def fastq_meta = []
+    if (!file(row.fastq_1).exists()) {
+        exit 1, "ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}"
+    }
+    if (meta.single_end) {
+        fastq_meta = [ meta, [ file(row.fastq_1) ] ]
+    } else {
+        if (!file(row.fastq_2).exists()) {
+            exit 1, "ERROR: Please check input samplesheet -> Read 2 FastQ file does not exist!\n${row.fastq_2}"
+        }
+        fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ]
+    }
+    return fastq_meta
+}
diff --git a/tower.yml b/tower.yml
new file mode 100644
index 00000000..787aedfe
--- /dev/null
+++ b/tower.yml
@@ -0,0 +1,5 @@
+reports:
+  multiqc_report.html:
+    display: "MultiQC HTML report"
+  samplesheet.csv:
+    display: "Auto-created samplesheet with collated metadata and FASTQ paths"
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
new file mode 100644
index 00000000..8d0e6008
--- /dev/null
+++ b/workflows/rnadeseq.nf
@@ -0,0 +1,133 @@
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    PRINT PARAMS SUMMARY
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation'
+
+def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs)
+def citation = '\n' + WorkflowMain.citation(workflow) + '\n'
+def summary_params = paramsSummaryMap(workflow)
+
+// Print parameter summary log to screen
+log.info logo + paramsSummaryLog(workflow) + citation
+
+WorkflowRnadeseq.initialise(params, log)
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    CONFIG FILES
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+ch_multiqc_config          = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true)
+ch_multiqc_custom_config   = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty()
+ch_multiqc_logo            = params.multiqc_logo   ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty()
+ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true)
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    IMPORT LOCAL MODULES/SUBWORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// SUBWORKFLOW: Consisting of a mix of local and nf-core/modules
+//
+include { INPUT_CHECK } from '../subworkflows/local/input_check'
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    IMPORT NF-CORE MODULES/SUBWORKFLOWS
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+//
+// MODULE: Installed directly from nf-core/modules
+//
+include { FASTQC                      } from '../modules/nf-core/fastqc/main'
+include { MULTIQC                     } from '../modules/nf-core/multiqc/main'
+include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main'
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    RUN MAIN WORKFLOW
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+// Info required for completion email and summary
+def multiqc_report = []
+
+workflow RNADESEQ {
+
+    ch_versions = Channel.empty()
+
+    //
+    // SUBWORKFLOW: Read in samplesheet, validate and stage input files
+    //
+    INPUT_CHECK (
+        file(params.input)
+    )
+    ch_versions = ch_versions.mix(INPUT_CHECK.out.versions)
+    // TODO: OPTIONAL, you can use nf-validation plugin to create an input channel from the samplesheet with Channel.fromSamplesheet("input")
+    // See the documentation https://nextflow-io.github.io/nf-validation/samplesheets/fromSamplesheet/
+    // ! There is currently no tooling to help you write a sample sheet schema
+
+    //
+    // MODULE: Run FastQC
+    //
+    FASTQC (
+        INPUT_CHECK.out.reads
+    )
+    ch_versions = ch_versions.mix(FASTQC.out.versions.first())
+
+    CUSTOM_DUMPSOFTWAREVERSIONS (
+        ch_versions.unique().collectFile(name: 'collated_versions.yml')
+    )
+
+    //
+    // MODULE: MultiQC
+    //
+    workflow_summary    = WorkflowRnadeseq.paramsSummaryMultiqc(workflow, summary_params)
+    ch_workflow_summary = Channel.value(workflow_summary)
+
+    methods_description    = WorkflowRnadeseq.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description, params)
+    ch_methods_description = Channel.value(methods_description)
+
+    ch_multiqc_files = Channel.empty()
+    ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml'))
+    ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml'))
+    ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect())
+    ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([]))
+
+    MULTIQC (
+        ch_multiqc_files.collect(),
+        ch_multiqc_config.toList(),
+        ch_multiqc_custom_config.toList(),
+        ch_multiqc_logo.toList()
+    )
+    multiqc_report = MULTIQC.out.report.toList()
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    COMPLETION EMAIL AND SUMMARY
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/
+
+workflow.onComplete {
+    if (params.email || params.email_on_fail) {
+        NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report)
+    }
+    NfcoreTemplate.summary(workflow, params, log)
+    if (params.hook_url) {
+        NfcoreTemplate.IM_notification(workflow, params, summary_params, projectDir, log)
+    }
+}
+
+/*
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+    THE END
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
+*/

From bb36b6f2de43d82a82b7adb7fbe58d5e30a41044 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 14 Jul 2023 15:37:36 +0200
Subject: [PATCH 074/124] linting

---
 assets/multiqc_config.yml              |  14 ++++++++------
 assets/nf-core-rnadeseq_logo_light.png | Bin 11674 -> 75779 bytes
 lib/NfcoreTemplate.groovy              |   5 +++++
 3 files changed, 13 insertions(+), 6 deletions(-)

diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index ee6b0b9e..b09e11cb 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -1,11 +1,13 @@
 report_comment: >
-  This report has been generated by the <a href="https://github.com/qbic-pipelines/rnadeseq" target="_blank">qbic-pipelines/rnadeseq</a>
-  analysis pipeline. For information about how to interpret these results, please see the
-  <a href="https://github.com/qbic-pipelines/rnadeseq" target="_blank">documentation</a>.
+  This report has been generated by the <a href="https://github.com/{{ name }}/{{ version}}" target="_blank">{{ name }}</a>
+  analysis pipeline.{% if branded %} For information about how to interpret these results, please see the
+  <a href="https://nf-co.re/{{ short_name }}/{{ version }}/output" target="_blank">documentation</a>.{% endif %}
 report_section_order:
-  software_versions:
+  "{{ name_noslash }}-methods-description":
     order: -1000
-  "qbic-pipelines-rnadeseq-summary":
+  software_versions:
     order: -1001
+  "{{ name_noslash }}-summary":
+    order: -1002
 
-export_plots: true
+export_plots: true
\ No newline at end of file
diff --git a/assets/nf-core-rnadeseq_logo_light.png b/assets/nf-core-rnadeseq_logo_light.png
index 007bdf2cf492bf8df85492075e6a1cd444ac52d2..f910229fda44f6e66f4cb00d92d8f4ed06721a47 100644
GIT binary patch
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diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
index 000bb929..408951ae 100755
--- a/lib/NfcoreTemplate.groovy
+++ b/lib/NfcoreTemplate.groovy
@@ -323,6 +323,11 @@ class NfcoreTemplate {
         String.format(
             """\n
             ${dashedLine(monochrome_logs)}
+                                                    ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset}
+            ${colors.blue}        ___     __   __   __   ___     ${colors.green}/,-._.--~\'${colors.reset}
+            ${colors.blue}  |\\ | |__  __ /  ` /  \\ |__) |__         ${colors.yellow}}  {${colors.reset}
+            ${colors.blue}  | \\| |       \\__, \\__/ |  \\ |___     ${colors.green}\\`-._,-`-,${colors.reset}
+                                                    ${colors.green}`._,._,\'${colors.reset}
             ${colors.purple}  ${workflow.manifest.name} ${workflow_version}${colors.reset}
             ${dashedLine(monochrome_logs)}
             """.stripIndent()

From 2df1239935d0e4fad3fb285514967a74b7b3c3dd Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 08:45:49 +0200
Subject: [PATCH 075/124] changed round_DE param to int

---
 assets/RNAseq_report.Rmd  | 10 +++---
 assets/multiqc_config.yml | 10 +++---
 bin/Execute_report.R      |  2 +-
 docs/usage.md             |  2 +-
 modules/local/report.nf   |  3 +-
 nextflow.config           |  3 +-
 nextflow_schema.json      | 75 +++++++++++++++++++++++++++++++--------
 7 files changed, 76 insertions(+), 29 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 67c6201f..a99df5f5 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1168,7 +1168,7 @@ if (isProvided(params$path_contrast_matrix)){
         DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
-        if (isProvided(params$round_DE)) {
+        if (isProvided(params$round_DE) && params$round_DE>-1) {
             DE_genes_contrast <- (DE_genes_contrast  %>% mutate_if(is.numeric, round, digits=params$round_DE))
         }
         # Save table
@@ -1228,7 +1228,7 @@ if (isProvided(params$path_contrast_list)) {
         DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
-        if (isProvided(params$round_DE)) {
+        if (isProvided(params$round_DE) && params$round_DE>-1) {
             DE_genes_contrast <- (DE_genes_contrast  %>% mutate_if(is.numeric, round, digits=params$round_DE))
         }
         # Save table
@@ -1271,7 +1271,7 @@ if (isProvided(params$path_contrast_pairs)) {
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
 
         # Round table before writing
-        if (isProvided(params$round_DE)) {
+        if (isProvided(params$round_DE) && params$round_DE>-1) {
             DE_genes_contrast <- (DE_genes_contrast  %>% mutate_if(is.numeric, round, digits=params$round_DE))
         }
 
@@ -1310,7 +1310,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
-        if (isProvided(params$round_DE)) {
+        if (isProvided(params$round_DE) && params$round_DE>-1) {
             DE_genes_contrast <- (DE_genes_contrast  %>% mutate_if(is.numeric, round, digits=params$round_DE))
         }
         # Save table
@@ -1373,7 +1373,7 @@ DE_genes_final_table = merge(x=DE_genes_final_table, y=gene_names, by.x="Ensembl
 DE_genes_final_table = DE_genes_final_table[,c(dim(DE_genes_final_table)[2],1:dim(DE_genes_final_table)[2]-1)]
 DE_genes_final_table = DE_genes_final_table[order(DE_genes_final_table[,"Ensembl_ID"]),]
 # Round table before writing
-if (isProvided(params$round_DE)) {
+if (isProvided(params$round_DE) && params$round_DE>-1) {
     DE_genes_final_table <- (DE_genes_final_table  %>% mutate_if(is.numeric, round, digits=params$round_DE))
 }
 
diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index b09e11cb..ccfd5b7e 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -1,13 +1,13 @@
 report_comment: >
-  This report has been generated by the <a href="https://github.com/{{ name }}/{{ version}}" target="_blank">{{ name }}</a>
-  analysis pipeline.{% if branded %} For information about how to interpret these results, please see the
-  <a href="https://nf-co.re/{{ short_name }}/{{ version }}/output" target="_blank">documentation</a>.{% endif %}
+  This report has been generated by the <a href="https://github.com/qbic-pipelines/rnadeseq" target="_blank">nf-core/crisprseq</a>
+  analysis pipeline. For information about how to interpret these results, please see the
+  <a href="https://github.com/qbic-pipelines/rnadeseq/blob/master/docs/output.md" target="_blank">documentation</a>.
 report_section_order:
-  "{{ name_noslash }}-methods-description":
+  "qbic-pipelines-rnadeseq-methods-description":
     order: -1000
   software_versions:
     order: -1001
-  "{{ name_noslash }}-summary":
+  "nf-core-rnadeseq-summary":
     order: -1002
 
 export_plots: true
\ No newline at end of file
diff --git a/bin/Execute_report.R b/bin/Execute_report.R
index c0463125..5b9160d1 100755
--- a/bin/Execute_report.R
+++ b/bin/Execute_report.R
@@ -21,7 +21,7 @@ option_list = list(
     make_option(c("-b", "--batch_effect"), action="store_true", default=FALSE, help="Batch effect correction."),
     make_option(c("-f", "--logFC_threshold"), type="double", default=NULL, help="Log Fold Change threshold to consider a gene DE."),
     make_option("--adj_pval_threshold", type="double", default=0.05, help="adjusted p value threshold to consider a gene DE."),
-    make_option("--round_DE", type="integer", default=NULL, help="How many decimals to keep after rounding the DE analysis values."),
+    make_option("--round_DE", type="integer", default=-1, help="How many decimals to keep after rounding the DE analysis values; if -1, will not round."),
 
     make_option(c("-z", "--norm_method"), type="character", default=NULL, help="Which transformation(s) to use."),
     make_option(c("-n", "--nsub_genes"), type="integer", default=NULL, help="Subset number of genes for vst."),
diff --git a/docs/usage.md b/docs/usage.md
index 0888e45b..13587b1f 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -336,7 +336,7 @@ This is ignored if `--norm_method` is set to `rlog`. If using the `vst` transfor
 
 ### `--round_DE`
 
-Integer indicating to how many decimals to round the DE results (default: no rounding).
+Integer indicating to how many decimals to round the DE results (default: -1, indicating no rounding).
 
 ### `--run_pathway_analysis`
 
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 8a4e235f..167fceb5 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -32,7 +32,6 @@ process REPORT {
     def genelist_opt = genelist.name != 'NO_FILE' ? "--genelist $genelist" : ''
     def relevel_opt = relevel.name != 'NO_FILE2' ? "--relevel $relevel" : ''
     def batch_effect_opt = params.batch_effect ? "--batch_effect TRUE" : ''
-    def round_DE_opt = params.round_DE ? "--round_DE $params.round_DE" : ''
 
     def pathway_opt = params.run_pathway_analysis ? "--pathway_analysis" : ''
     def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : ''
@@ -68,7 +67,7 @@ process REPORT {
         --adj_pval_threshold $params.adj_pval_threshold \
         --norm_method $params.norm_method \
         --nsub_genes $params.vst_genes_number \
-        $round_DE_opt \
+        --round_DE $params.round_DE \
         $pathway_opt \
         $custom_gmt_opt \
         $set_background_opt \
diff --git a/nextflow.config b/nextflow.config
index 81eb4456..f9b783d2 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -28,7 +28,7 @@ params {
     adj_pval_threshold = 0.05
     norm_method = 'rlog'
     vst_genes_number = 1000
-    round_DE = null
+    round_DE = -1
 
     // Pathway analysis args
     //* organism = null
@@ -60,6 +60,7 @@ params {
 
     // Boilerplate options
     email = null
+    email_on_fail = null
     plaintext_email = false
     monochrome_logs = false
     help = false
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 8c7b2e1a..0823a8c6 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -32,7 +32,12 @@
                     "description": "Not necessary if genome is set or input_type is featurecounts. GTF file to be used for DESeq if input is rsem or salmon."
                 }
             },
-            "required": ["input_type", "gene_counts", "input", "model"]
+            "required": [
+                "input_type",
+                "gene_counts",
+                "input",
+                "model"
+            ]
         },
         "deseq2_arguments": {
             "title": "DESeq2 arguments",
@@ -83,7 +88,11 @@
                 "norm_method": {
                     "type": "string",
                     "default": "rlog",
-                    "enum": ["rlog", "vst", "vst-force"],
+                    "enum": [
+                        "rlog",
+                        "vst",
+                        "vst-force"
+                    ],
                     "description": "Which transformation to use during DE analysis; either rlog or vst or vst-force . If vst: the pipeline will override this and run rlog if it detects a strong variation in size factors in the input data (this will be stated in the report). To disable this behavior, set to vst-force."
                 },
                 "vst_genes_number": {
@@ -92,9 +101,9 @@
                     "description": "Integer indicating how many genes to subset to during vst transformation (when using rlog, will ignore this number). Default: 1000."
                 },
                 "round_DE": {
-                    "type": "string",
-                    "default": "null",
-                    "description": "Number of decimals to which to round result tables, default: no rounding. Integer, this is only set to string in the schema to allow for a null default."
+                    "type": "integer",
+                    "default": -1,
+                    "description": "Number of decimals to which to round result tables, default: -1 (this means no rounding)."
                 }
             }
         },
@@ -118,7 +127,6 @@
                 },
                 "run_pathway_analysis": {
                     "type": "boolean",
-                    "default": false,
                     "description": "Turn on this flag if you wish to run pathway analysis."
                 },
                 "custom_gmt": {
@@ -184,7 +192,6 @@
                 },
                 "igenomes_ignore": {
                     "type": "boolean",
-                    "default": false,
                     "description": "Do not load the iGenomes reference config."
                 }
             }
@@ -197,7 +204,6 @@
             "properties": {
                 "citest": {
                     "type": "boolean",
-                    "default": false,
                     "description": "Only enable for CI tests on the github servers",
                     "hidden": true
                 }
@@ -215,7 +221,14 @@
                     "description": "Method used to save pipeline results to output directory.",
                     "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.",
                     "fa_icon": "fas fa-copy",
-                    "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"],
+                    "enum": [
+                        "symlink",
+                        "rellink",
+                        "link",
+                        "copy",
+                        "copyNoFollow",
+                        "move"
+                    ],
                     "hidden": true
                 },
                 "outdir": {
@@ -249,7 +262,6 @@
                     "type": "boolean",
                     "description": "Send plain-text email instead of HTML.",
                     "fa_icon": "fas fa-remove-format",
-                    "default": false,
                     "hidden": true
                 },
                 "monochrome_logs": {
@@ -316,7 +328,6 @@
                     "type": "boolean",
                     "fa_icon": "far fa-eye-slash",
                     "description": "Show all params when using `--help`",
-                    "default": false,
                     "hidden": true,
                     "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters."
                 },
@@ -324,7 +335,6 @@
                     "type": "boolean",
                     "fa_icon": "far fa-check-circle",
                     "description": "Validation of parameters fails when an unrecognised parameter is found.",
-                    "default": false,
                     "hidden": true,
                     "help_text": "By default, when an unrecognised parameter is found, it returns a warinig."
                 },
@@ -332,11 +342,45 @@
                     "type": "boolean",
                     "fa_icon": "far fa-check-circle",
                     "description": "Validation of parameters in lenient more.",
-                    "default": false,
                     "hidden": true,
                     "help_text": "Allows string values that are parseable as numbers or booleans. For further information see [JSONSchema docs](https://github.com/everit-org/json-schema#lenient-mode)."
                 }
             }
+        },
+        "max_job_request_options": {
+            "title": "Max job request options",
+            "type": "object",
+            "fa_icon": "fab fa-acquisitions-incorporated",
+            "description": "Set the top limit for requested resources for any single job.",
+            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
+            "properties": {
+                "max_cpus": {
+                    "type": "integer",
+                    "description": "Maximum number of CPUs that can be requested for any single job.",
+                    "default": 16,
+                    "fa_icon": "fas fa-microchip",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`"
+                },
+                "max_memory": {
+                    "type": "string",
+                    "description": "Maximum amount of memory that can be requested for any single job.",
+                    "default": "128.GB",
+                    "fa_icon": "fas fa-memory",
+                    "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`"
+                },
+                "max_time": {
+                    "type": "string",
+                    "description": "Maximum amount of time that can be requested for any single job.",
+                    "default": "240.h",
+                    "fa_icon": "far fa-clock",
+                    "pattern": "^(\\d+\\.?\\s*(s|m|h|d|day)\\s*)+$",
+                    "hidden": true,
+                    "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`"
+                }
+            }
         }
     },
     "allOf": [
@@ -363,6 +407,9 @@
         },
         {
             "$ref": "#/definitions/boilerplate_options"
+        },
+        {
+            "$ref": "#/definitions/max_job_request_options"
         }
     ]
-}
+}
\ No newline at end of file

From 7b3dfc260f8063fbeb06158fbbbbeaf4a22f128f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 08:48:23 +0200
Subject: [PATCH 076/124] Updated Changelog

---
 CHANGELOG.md | 1 +
 1 file changed, 1 insertion(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 93201a2e..b81e816a 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#209](https://github.com/qbic-pipelines/rnadeseq/pull/209) Template update to 2.9, Chromium Falcon; exchanged file.exists for nf-core validation checks; changed round_DE param to int
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_library_scaled_gene_counts.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
 - [#205](https://github.com/qbic-pipelines/rnadeseq/pull/205) Template update
 - [#204](https://github.com/qbic-pipelines/rnadeseq/pull/204) Changed relevel path in test_relevel.config to the qbic-pipelines repo

From 6a788aba7b2f71deb75ac65758b5462e7b4092e4 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 09:11:40 +0200
Subject: [PATCH 077/124] prettier, updated ci nextflow version

---
 .github/workflows/ci.yml  |  2 +-
 assets/multiqc_config.yml |  2 +-
 nextflow_schema.json      | 24 ++++--------------------
 3 files changed, 6 insertions(+), 22 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index b271ef6e..6e833767 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -60,7 +60,7 @@ jobs:
     name: Test workflow parameters
     runs-on: ubuntu-latest
     env:
-      NXF_VER: "22.10.1"
+      NXF_VER: "23.04.0"
       NXF_ANSI_LOG: false
     strategy:
       # This tells github to continue running other profile tests if some tests fail (default cancels all tests upon failure)
diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index ccfd5b7e..c5414993 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -10,4 +10,4 @@ report_section_order:
   "nf-core-rnadeseq-summary":
     order: -1002
 
-export_plots: true
\ No newline at end of file
+export_plots: true
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 0823a8c6..7820fe73 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -32,12 +32,7 @@
                     "description": "Not necessary if genome is set or input_type is featurecounts. GTF file to be used for DESeq if input is rsem or salmon."
                 }
             },
-            "required": [
-                "input_type",
-                "gene_counts",
-                "input",
-                "model"
-            ]
+            "required": ["input_type", "gene_counts", "input", "model"]
         },
         "deseq2_arguments": {
             "title": "DESeq2 arguments",
@@ -88,11 +83,7 @@
                 "norm_method": {
                     "type": "string",
                     "default": "rlog",
-                    "enum": [
-                        "rlog",
-                        "vst",
-                        "vst-force"
-                    ],
+                    "enum": ["rlog", "vst", "vst-force"],
                     "description": "Which transformation to use during DE analysis; either rlog or vst or vst-force . If vst: the pipeline will override this and run rlog if it detects a strong variation in size factors in the input data (this will be stated in the report). To disable this behavior, set to vst-force."
                 },
                 "vst_genes_number": {
@@ -221,14 +212,7 @@
                     "description": "Method used to save pipeline results to output directory.",
                     "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.",
                     "fa_icon": "fas fa-copy",
-                    "enum": [
-                        "symlink",
-                        "rellink",
-                        "link",
-                        "copy",
-                        "copyNoFollow",
-                        "move"
-                    ],
+                    "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"],
                     "hidden": true
                 },
                 "outdir": {
@@ -412,4 +396,4 @@
             "$ref": "#/definitions/max_job_request_options"
         }
     ]
-}
\ No newline at end of file
+}

From 8b9b9c77962dcda6b9d2987f4f2bf617211899d4 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 09:19:57 +0200
Subject: [PATCH 078/124] corrected multiqc_config, changed batch_effect to
 boolean

---
 assets/multiqc_config.yml | 4 ++--
 nextflow.config           | 2 +-
 nextflow_schema.json      | 2 +-
 3 files changed, 4 insertions(+), 4 deletions(-)

diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index c5414993..78eaf597 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -1,5 +1,5 @@
 report_comment: >
-  This report has been generated by the <a href="https://github.com/qbic-pipelines/rnadeseq" target="_blank">nf-core/crisprseq</a>
+  This report has been generated by the <a href="https://github.com/qbic-pipelines/rnadeseq" target="_blank">qbic-pipelines/rnadeseq</a>
   analysis pipeline. For information about how to interpret these results, please see the
   <a href="https://github.com/qbic-pipelines/rnadeseq/blob/master/docs/output.md" target="_blank">documentation</a>.
 report_section_order:
@@ -7,7 +7,7 @@ report_section_order:
     order: -1000
   software_versions:
     order: -1001
-  "nf-core-rnadeseq-summary":
+  "nf-core-rnadeseq-summary": # this is necessary to prevent a linting error
     order: -1002
 
 export_plots: true
diff --git a/nextflow.config b/nextflow.config
index f9b783d2..2c01bd7b 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -23,7 +23,7 @@ params {
     contrast_pairs = 'DEFAULT2'
     genelist = 'NO_FILE'
     relevel = 'NO_FILE2'
-    batch_effect = null
+    batch_effect = false
     logFC_threshold = 0
     adj_pval_threshold = 0.05
     norm_method = 'rlog'
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 7820fe73..1acdf285 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -66,7 +66,7 @@
                     "description": "Tsv indicating list of factors (conditions in the metadata table/samplesheet) and the new level on which to relevel the factor. Check contrasts docs."
                 },
                 "batch_effect": {
-                    "type": "string",
+                    "type": "boolean",
                     "default": "false",
                     "description": "Turn on this flag if you wish to consider batch effects. You need to add the batch effect to the linear model too!"
                 },

From adf5db97e323468545e19d54d9533eb076ec6787 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 09:45:45 +0200
Subject: [PATCH 079/124] changed some occurrences of nf-core/rnadeseq to
 qbic-pipelines/rnadeseq, removed samplesheet.csv from assets

---
 .github/CONTRIBUTING.md                    | 10 +++++-----
 .github/ISSUE_TEMPLATE/bug_report.yml      |  2 +-
 .github/ISSUE_TEMPLATE/config.yml          |  2 +-
 .github/ISSUE_TEMPLATE/feature_request.yml |  2 +-
 .github/PULL_REQUEST_TEMPLATE.md           |  1 -
 assets/samplesheet.csv                     |  3 ---
 assets/schema_input.json                   |  4 ++--
 lib/WorkflowMain.groovy                    |  2 +-
 lib/WorkflowRnadeseq.groovy                |  2 +-
 modules/local/samplesheet_check.nf         |  2 +-
 nextflow.config                            |  4 ++--
 nextflow_schema.json                       |  1 -
 12 files changed, 15 insertions(+), 20 deletions(-)
 delete mode 100644 assets/samplesheet.csv

diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md
index 3cc63d1d..1ea3fb25 100644
--- a/.github/CONTRIBUTING.md
+++ b/.github/CONTRIBUTING.md
@@ -1,9 +1,9 @@
 # qbic-pipelines/rnadeseq: Contributing Guidelines
 
 Hi there!
-Many thanks for taking an interest in improving nf-core/rnadeseq.
+Many thanks for taking an interest in improving qbic-pipelines/rnadeseq.
 
-We try to manage the required tasks for nf-core/rnadeseq using GitHub issues, you probably came to this page when creating one.
+We try to manage the required tasks for qbic-pipelines/rnadeseq using GitHub issues, you probably came to this page when creating one.
 Please use the pre-filled template to save time.
 
 However, don't be put off by this template - other more general issues and suggestions are welcome!
@@ -13,8 +13,8 @@ Contributions to the code are even more welcome ;)
 
 If you'd like to write some code for qbic-pipelines/rnadeseq, the standard workflow is as follows:
 
-1. Check that there isn't already an issue about your idea in the [nf-core/rnadeseq issues](https://github.com/nf-core/rnadeseq/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this
-2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/rnadeseq repository](https://github.com/nf-core/rnadeseq) to your GitHub account
+1. Check that there isn't already an issue about your idea in the [qbic-pipelines/rnadeseq issues](https://github.com/qbic-pipelines/rnadeseq/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this
+2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [qbic-pipelines/rnadeseq repository](https://github.com/qbic-pipelines/rnadeseq) to your GitHub account
 3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions)
 4. Use `nf-core schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10).
 5. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged
@@ -106,7 +106,7 @@ This repo includes a devcontainer configuration which will create a GitHub Codes
 
 To get started:
 
-- Open the repo in [Codespaces](https://github.com/nf-core/rnadeseq/codespaces)
+- Open the repo in [Codespaces](https://github.com/qbic-pipelines/rnadeseq/codespaces)
 - Tools installed
   - nf-core
   - Nextflow
diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml
index 15accb05..6aa7ac17 100644
--- a/.github/ISSUE_TEMPLATE/bug_report.yml
+++ b/.github/ISSUE_TEMPLATE/bug_report.yml
@@ -47,4 +47,4 @@ body:
         * Executor _(eg. slurm, local, awsbatch)_
         * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, or Apptainer)_
         * OS _(eg. CentOS Linux, macOS, Linux Mint)_
-        * Version of nf-core/rnadeseq _(eg. 1.1, 1.5, 1.8.2)_
+        * Version of qbic-pipelines/rnadeseq _(eg. 1.1, 1.5, 1.8.2)_
diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml
index 1c032cae..a5de823c 100644
--- a/.github/ISSUE_TEMPLATE/config.yml
+++ b/.github/ISSUE_TEMPLATE/config.yml
@@ -4,4 +4,4 @@ contact_links:
     about: Please join the nf-core community here
   - name: "Slack #rnadeseq channel"
     url: https://nfcore.slack.com/channels/rnadeseq
-    about: Discussion about the nf-core/rnadeseq pipeline
+    about: Discussion about the qbic-pipelines/rnadeseq pipeline
diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml
index 6bd51f6c..abf60a68 100644
--- a/.github/ISSUE_TEMPLATE/feature_request.yml
+++ b/.github/ISSUE_TEMPLATE/feature_request.yml
@@ -1,5 +1,5 @@
 name: Feature request
-description: Suggest an idea for the nf-core/rnadeseq pipeline
+description: Suggest an idea for the qbic-pipelines/rnadeseq pipeline
 labels: enhancement
 body:
   - type: textarea
diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md
index b4ccca5e..fd829b68 100644
--- a/.github/PULL_REQUEST_TEMPLATE.md
+++ b/.github/PULL_REQUEST_TEMPLATE.md
@@ -16,7 +16,6 @@ Learn more about contributing: [CONTRIBUTING.md](https://github.com/qbic-pipelin
 - [ ] This comment contains a description of changes (with reason).
 - [ ] If you've fixed a bug or added code that should be tested, add tests!
 - [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/qbic-pipelines/rnadeseq/tree/master/.github/CONTRIBUTING.md)
-- [ ] If necessary, also make a PR on the nf-core/rnadeseq _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository.
 - [ ] Make sure your code lints (`nf-core lint`).
 - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir <OUTDIR>`).
 - [ ] Usage Documentation in `docs/usage.md` is updated.
diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv
deleted file mode 100644
index 5f653ab7..00000000
--- a/assets/samplesheet.csv
+++ /dev/null
@@ -1,3 +0,0 @@
-sample,fastq_1,fastq_2
-SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz
-SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz,
diff --git a/assets/schema_input.json b/assets/schema_input.json
index bd54c145..8f83ef7d 100644
--- a/assets/schema_input.json
+++ b/assets/schema_input.json
@@ -1,7 +1,7 @@
 {
     "$schema": "http://json-schema.org/draft-07/schema",
-    "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/assets/schema_input.json",
-    "title": "nf-core/rnadeseq pipeline - params.input schema",
+    "$id": "https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/master/assets/schema_input.json",
+    "title": "qbic-pipelines/rnadeseq pipeline - params.input schema",
     "description": "Schema for the file provided with params.input",
     "type": "array",
     "items": {
diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy
index f6bcde2c..3ae24dbf 100755
--- a/lib/WorkflowMain.groovy
+++ b/lib/WorkflowMain.groovy
@@ -1,5 +1,5 @@
 //
-// This file holds several functions specific to the main.nf workflow in the nf-core/rnadeseq pipeline
+// This file holds several functions specific to the main.nf workflow in the qbic-pipelines/rnadeseq pipeline
 //
 
 import nextflow.Nextflow
diff --git a/lib/WorkflowRnadeseq.groovy b/lib/WorkflowRnadeseq.groovy
index dc726df4..7a1ae2bd 100755
--- a/lib/WorkflowRnadeseq.groovy
+++ b/lib/WorkflowRnadeseq.groovy
@@ -1,5 +1,5 @@
 //
-// This file holds several functions specific to the workflow/rnadeseq.nf in the nf-core/rnadeseq pipeline
+// This file holds several functions specific to the workflow/rnadeseq.nf in the qbic-pipelines/rnadeseq pipeline
 //
 
 import nextflow.Nextflow
diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf
index c1d02d25..d3ebb676 100644
--- a/modules/local/samplesheet_check.nf
+++ b/modules/local/samplesheet_check.nf
@@ -17,7 +17,7 @@ process SAMPLESHEET_CHECK {
     when:
     task.ext.when == null || task.ext.when
 
-    script: // This script is bundled with the pipeline, in nf-core/rnadeseq/bin/
+    script: // This script is bundled with the pipeline, in qbic-pipelines/rnadeseq/bin/
     """
     check_samplesheet.py \\
         $samplesheet \\
diff --git a/nextflow.config b/nextflow.config
index 2c01bd7b..fd1595d6 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -245,9 +245,9 @@ dag {
 }
 
 manifest {
-    name            = 'nf-core/rnadeseq'
+    name            = 'qbic-pipelines/rnadeseq'
     author          = """Gisela Gabernet, Silvia Morini, Oskar Wacker, QBiC"""
-    homePage        = 'https://github.com/nf-core/rnadeseq'
+    homePage        = 'https://github.com/qbic-pipelines/rnadeseq'
     description     = """Differential gene expression analysis and pathway analysis of RNAseq data"""
     mainScript      = 'main.nf'
     nextflowVersion = '!>=23.04.0'
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 1acdf285..866131e2 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -67,7 +67,6 @@
                 },
                 "batch_effect": {
                     "type": "boolean",
-                    "default": "false",
                     "description": "Turn on this flag if you wish to consider batch effects. You need to add the batch effect to the linear model too!"
                 },
                 "logFC_threshold": {

From f5fbbae24974cda357a4fa237d446649ea96eb76 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 10:01:38 +0200
Subject: [PATCH 080/124] lint

---
 assets/multiqc_config.yml | 2 +-
 lib/NfcoreTemplate.groovy | 5 -----
 2 files changed, 1 insertion(+), 6 deletions(-)

diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index 78eaf597..83ebb310 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -7,7 +7,7 @@ report_section_order:
     order: -1000
   software_versions:
     order: -1001
-  "nf-core-rnadeseq-summary": # this is necessary to prevent a linting error
+  "qbic-pipelines-rnadeseq-summary": # this is necessary to prevent a linting error
     order: -1002
 
 export_plots: true
diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy
index 408951ae..000bb929 100755
--- a/lib/NfcoreTemplate.groovy
+++ b/lib/NfcoreTemplate.groovy
@@ -323,11 +323,6 @@ class NfcoreTemplate {
         String.format(
             """\n
             ${dashedLine(monochrome_logs)}
-                                                    ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset}
-            ${colors.blue}        ___     __   __   __   ___     ${colors.green}/,-._.--~\'${colors.reset}
-            ${colors.blue}  |\\ | |__  __ /  ` /  \\ |__) |__         ${colors.yellow}}  {${colors.reset}
-            ${colors.blue}  | \\| |       \\__, \\__/ |  \\ |___     ${colors.green}\\`-._,-`-,${colors.reset}
-                                                    ${colors.green}`._,._,\'${colors.reset}
             ${colors.purple}  ${workflow.manifest.name} ${workflow_version}${colors.reset}
             ${dashedLine(monochrome_logs)}
             """.stripIndent()

From d65af8348b15e03b9a7fb94d3977e44f21d46dca Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 11:49:01 +0200
Subject: [PATCH 081/124] added file exists checks to schema, reordered
 mqc_config

---
 assets/multiqc_config.yml | 4 ++--
 nextflow_schema.json      | 6 ++++++
 2 files changed, 8 insertions(+), 2 deletions(-)

diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml
index 83ebb310..737c3d27 100644
--- a/assets/multiqc_config.yml
+++ b/assets/multiqc_config.yml
@@ -3,11 +3,11 @@ report_comment: >
   analysis pipeline. For information about how to interpret these results, please see the
   <a href="https://github.com/qbic-pipelines/rnadeseq/blob/master/docs/output.md" target="_blank">documentation</a>.
 report_section_order:
-  "qbic-pipelines-rnadeseq-methods-description":
-    order: -1000
   software_versions:
     order: -1001
   "qbic-pipelines-rnadeseq-summary": # this is necessary to prevent a linting error
     order: -1002
+  "qbic-pipelines-rnadeseq-methods-description":
+    order: -1000
 
 export_plots: true
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 866131e2..8fb91596 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -17,14 +17,20 @@
                 },
                 "gene_counts": {
                     "type": "string",
+                    "format": "file-path",
+                    "exists": true,
                     "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder."
                 },
                 "input": {
                     "type": "string",
+                    "format": "file-path",
+                    "exists": true,
                     "description": "Metadata table/samplesheet (TSV). Rows are samples and columns contain sample grouping."
                 },
                 "model": {
                     "type": "string",
+                    "format": "file-path",
+                    "exists": true,
                     "description": "Linear model function to calculate the contrasts (TXT). Variable names should be columns in metadata file/samplesheet."
                 },
                 "gtf": {

From 2fe121b760a165875200b82f08952c96ead1c9d9 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 17 Jul 2023 11:54:58 +0200
Subject: [PATCH 082/124] removed exists check for gene_counts as this can also
 be a dir instead of file

---
 nextflow_schema.json | 2 --
 1 file changed, 2 deletions(-)

diff --git a/nextflow_schema.json b/nextflow_schema.json
index 8fb91596..3aec2e6a 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -17,8 +17,6 @@
                 },
                 "gene_counts": {
                     "type": "string",
-                    "format": "file-path",
-                    "exists": true,
                     "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder."
                 },
                 "input": {

From 6d6b629d93454685d4dd050c792cee2939512f59 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 19 Jul 2023 16:04:05 +0200
Subject: [PATCH 083/124] removed kaleido, static heatmaps are now created with
 pheatmap

---
 .../workflows/push_github_container_reg.yml   |  4 ++--
 .nf-core.yml                                  |  1 -
 assets/RNAseq_report.Rmd                      | 21 ++++++++++++-------
 environment.yml                               |  4 +---
 4 files changed, 17 insertions(+), 13 deletions(-)

diff --git a/.github/workflows/push_github_container_reg.yml b/.github/workflows/push_github_container_reg.yml
index b8934ae3..abb3bb14 100644
--- a/.github/workflows/push_github_container_reg.yml
+++ b/.github/workflows/push_github_container_reg.yml
@@ -9,8 +9,8 @@ on:
     types: [published]
 
 jobs:
-  push_dockerhub:
-    name: Push new Docker image to Docker Hub
+  push_github:
+    name: Push new Docker image to GHCR
     runs-on: ubuntu-latest
     # Only run for the qbic-pipelines repo, for releases and merged PRs
     if: ${{ github.repository == 'qbic-pipelines/rnadeseq' }}
diff --git a/.nf-core.yml b/.nf-core.yml
index a797f271..6d79fbe1 100644
--- a/.nf-core.yml
+++ b/.nf-core.yml
@@ -20,7 +20,6 @@ lint:
     - .github/ISSUE_TEMPLATE/config.yml
     - .github/workflows/awsfulltest.yml
     - .github/workflows/awstest.yml
-    - .github/workflows/push_dockerhub.yml
     - assets/multiqc_config.yaml
     - assets/nf-core-qbic-pipelines/rnadeseq_logo_light.png
     - bin/markdown_to_html.r
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index a99df5f5..0bf7d692 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -220,9 +220,6 @@ invisible( lapply(c(
 "tools",
 "yaml"
 ), library, character.only=T))
-
-# Import python sys so that the save_image command works
-reticulate::py_run_string("import sys")
 ```
 
 ```{r create_outdirs, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
@@ -1065,14 +1062,24 @@ sampleDists <- dist(t(assay(if (run_rlog) rld else vsd)))
 sampleDistMatrix <- as.matrix(sampleDists)
 colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255)
 par(oma=c(3,3,3,3))
+dist_heatmap <- pheatmap(mat = sampleDistMatrix, color=colors)
+
+svg("differential_gene_expression/plots/Heatmaps_of_distances.svg")
+dist_heatmap
+dev.off()
+png("differential_gene_expression/plots/Heatmaps_of_distances.png")
+dist_heatmap
+dev.off()
+pdf("differential_gene_expression/plots/Heatmaps_of_distances.pdf")
+dist_heatmap
+dev.off()
+
+# Recompute heatmap with heatmaply for interactive plot in report
 dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey")
-save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.pdf")
-save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.png")
-save_image(dist_heatmap, file="differential_gene_expression/plots/Heatmaps_of_distances.svg")
 ```
 
 ```{r heatmap_show, echo=FALSE, message=FALSE, warning=FALSE, out.width="90%", out.height="130%", fig.asp=1, dpi=1000, fig.align='center'}
-#add export button to plot
+# Add export button to plot
 dist_heatmap %>% layout(height=700, width=800) %>% config(modeBarButtonsToAdd = list(svg_exp))
 ```
 
diff --git a/environment.yml b/environment.yml
index fe104828..3e62e0e1 100644
--- a/environment.yml
+++ b/environment.yml
@@ -22,7 +22,6 @@ dependencies:
   - bioconda::bioconductor-vsn=3.62.0
   - conda-forge::pandoc=2.17.1.1
   - conda-forge::plotly=5.13.0 #this is necessary for exporting plots with save_image()
-  - conda-forge::python-kaleido=0.2.1 #this is necessary for exporting plots with save_image()
   - conda-forge::r-base=4.1.2
   - conda-forge::r-details=0.3.0
   - conda-forge::r-dplyr=1.0.10
@@ -42,9 +41,8 @@ dependencies:
   - conda-forge::r-plyr=1.8.6
   - conda-forge::r-rcolorbrewer=1.1_2
   - conda-forge::r-reshape2=1.4.4
-  - conda-forge::r-reticulate=1.28 #this is necessary for exporting heatmaply plots with save_image()
   - conda-forge::r-rmarkdown=2.11
   - conda-forge::r-sessioninfo=1.2.2
   - conda-forge::r-svglite=2.1.0
   - conda-forge::r-tidyverse=1.3.1
-  - conda-forge::r-yaml=2.2.2
+  - conda-forge::r-yaml=2.2.2
\ No newline at end of file

From ff3d143e075d1eb0d447bcea0aa49e25f1c2eb31 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 19 Jul 2023 16:07:53 +0200
Subject: [PATCH 084/124] updated changelog

---
 CHANGELOG.md | 1 +
 1 file changed, 1 insertion(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index b81e816a..c6ef21ba 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#211](https://github.com/qbic-pipelines/rnadeseq/pull/21) Replaced heatmaply with pheatmap for static plots and removed kaleido and reticulate from container
 - [#209](https://github.com/qbic-pipelines/rnadeseq/pull/209) Template update to 2.9, Chromium Falcon; exchanged file.exists for nf-core validation checks; changed round_DE param to int
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_library_scaled_gene_counts.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
 - [#205](https://github.com/qbic-pipelines/rnadeseq/pull/205) Template update

From b17db5152fae2d77a2440b97dda2f05f8964eafa Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 20 Jul 2023 11:56:03 +0200
Subject: [PATCH 085/124] removed plotly from env

---
 assets/RNAseq_report.Rmd | 1 -
 environment.yml          | 3 +--
 2 files changed, 1 insertion(+), 3 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 0bf7d692..03ac0621 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -209,7 +209,6 @@ invisible( lapply(c(
 "kableExtra",
 "knitr",
 "formattable",
-"plotly",
 "webshot",
 "htmlwidgets",
 "pheatmap",
diff --git a/environment.yml b/environment.yml
index 3e62e0e1..3c62aa8a 100644
--- a/environment.yml
+++ b/environment.yml
@@ -21,7 +21,6 @@ dependencies:
   - bioconda::bioconductor-tximport=1.22.0
   - bioconda::bioconductor-vsn=3.62.0
   - conda-forge::pandoc=2.17.1.1
-  - conda-forge::plotly=5.13.0 #this is necessary for exporting plots with save_image()
   - conda-forge::r-base=4.1.2
   - conda-forge::r-details=0.3.0
   - conda-forge::r-dplyr=1.0.10
@@ -45,4 +44,4 @@ dependencies:
   - conda-forge::r-sessioninfo=1.2.2
   - conda-forge::r-svglite=2.1.0
   - conda-forge::r-tidyverse=1.3.1
-  - conda-forge::r-yaml=2.2.2
\ No newline at end of file
+  - conda-forge::r-yaml=2.2.2

From 1b4970d5f38c292b6431ee813c615cc929fa785d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 26 Jul 2023 10:24:18 +0200
Subject: [PATCH 086/124] fixed enrichment plots which were not changable in
 the report since I limited the y axis

---
 assets/RNAseq_report.Rmd | 5 ++++-
 1 file changed, 4 insertions(+), 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 03ac0621..63258414 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -2049,13 +2049,16 @@ if (length(q_list) > 0) {
     }
 
     path_enrich <- as.data.frame(gostres$result)
+
     if (nrow(path_enrich) > 0){
         pg2 <- gostplot(gostres, capped=T, interactive=T)
         pg2[['x']][['layout']][['annotations']][[1]][['x']] <- -params$adj_pval_threshold
 
         #limit gostplot y maximum
         for (counter in c(1:length(contrast_files))) {
-            pg2[['x']][['layout']][[paste0('yaxis', ifelse(counter>1, counter, ""))]][['range']] <- c(0,ceiling(2*-log10(max(gostres$p_value))))
+            expression <- parse(text=paste0("pg2 %>% layout(yaxis", ifelse(counter>1, counter, "")," = list(range = list(0, ceiling(2*-log10(max(gostres$p_value))))))"))
+            pg2 <- eval(expression)
+            pg2 %>% layout(yaxis2 = list(range = list(0, ceiling(2*-log10(max(gostres$p_value))))))
         }
 
         #add export button to plot

From 6e9ec341a71748cb8d26b99f0d224268e6179788 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 28 Jul 2023 17:05:47 +0200
Subject: [PATCH 087/124] added comp methods when no software_versions, fixed
 enrichment plot layout

---
 assets/RNAseq_report.Rmd | 23 +++++++++++++++++------
 1 file changed, 17 insertions(+), 6 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 63258414..0fe78896 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -2050,15 +2050,19 @@ if (length(q_list) > 0) {
 
     path_enrich <- as.data.frame(gostres$result)
 
+    # Determine which max has to be set for the gostplot y axis: highest -log10(pval) (use min function as the smallest
+    # p value produces the highest -log10) OR 17, as the gostplot caps at 16 (values>16 are set to 17)
+    y_max <- min(ceiling(-log10(min(path_enrich$p_value, na.rm=T))), 18)
+
     if (nrow(path_enrich) > 0){
         pg2 <- gostplot(gostres, capped=T, interactive=T)
         pg2[['x']][['layout']][['annotations']][[1]][['x']] <- -params$adj_pval_threshold
 
-        #limit gostplot y maximum
+        # limit gostplot y maximum dynamically for all subplots
         for (counter in c(1:length(contrast_files))) {
-            expression <- parse(text=paste0("pg2 %>% layout(yaxis", ifelse(counter>1, counter, "")," = list(range = list(0, ceiling(2*-log10(max(gostres$p_value))))))"))
+
+            expression <- parse(text=paste0("pg2 %>% layout(yaxis", ifelse(counter>1, counter, "")," = list(range = list(0, y_max)))"))
             pg2 <- eval(expression)
-            pg2 %>% layout(yaxis2 = list(range = list(0, ceiling(2*-log10(max(gostres$p_value))))))
         }
 
         #add export button to plot
@@ -2113,8 +2117,9 @@ quote_string <- ifelse(
 )
 cat(quote_string)
 
-# Initiate var in case no software_versions were provided
+# Initiate vars in case no software_versions were provided
 software_versions_csv <- F
+quant_tool <- params$input_type
 ```
 
 Further support for this project will be restricted to the results presented in this report.
@@ -2153,6 +2158,10 @@ if (software_versions_csv) {
 
 # Computational methods
 
+```{r no_software_versions, echo=FALSE, results='asis', eval=!isProvided(params$path_software_versions)}
+cat("No software versions file was provided for primary analysis.")
+```
+
 ```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
 
 # In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is obvious in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
@@ -2181,10 +2190,12 @@ quant_cite,
 ))
 
 ```{r MQC_cite, echo=FALSE, results='asis', eval=software_versions_csv&isProvided(params$path_software_versions)}
+
 cat(paste0("\nAn aggregation of the quality control for the RNAseq analysis was performed with `MultiQC ", as.character(version["MultiQC"]), "` [^2] [@ewels2016multiqc]."))
+
 ```
 
-```{r software_versions_DE, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
+```{r software_versions_DE, echo=FALSE, results='asis'}
 
 cat(paste0(
 "## Differential gene expression analysis
@@ -2199,7 +2210,7 @@ pval_text, " for the p-value and ",
 logFC_text, " for the log2 Fold Change."
 ))
 
-```{r conditional_pathway, echo=FALSE, results='asis', eval=isProvided(params$pathway_analysis)&isProvided(params$path_software_versions)}
+```{r conditional_pathway, echo=FALSE, results='asis', eval=isProvided(params$pathway_analysis)}
 version_gprofiler2 = packageVersion("gprofiler2")
 version_annotation = packageVersion("AnnotationDbi")
 name_species       = params$species_library

From 084a9877a2c43eb86ed0cc2e4c6fef28624e1cf6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 28 Jul 2023 17:11:50 +0200
Subject: [PATCH 088/124] updated Changelog

---
 CHANGELOG.md | 2 ++
 1 file changed, 2 insertions(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index c6ef21ba..2b5292b6 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
+- [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
 - [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
 - [#197](https://github.com/qbic-pipelines/rnadeseq/pull/197) Added gprofiler version string to report
@@ -25,6 +26,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Fixed
 
+- [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Fixed movability of interactive gostplots
 - [#208](https://github.com/qbic-pipelines/rnadeseq/pull/208) Fixed relevel bug, the function should now finally work!
 - [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added correct plot titles to meanSdPlot (depending on normalization)

From fdd93bc7605d14c9d2293b55aab48223b8d17bd6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 3 Aug 2023 13:24:50 +0200
Subject: [PATCH 089/124] working on smrnaseq import

---
 assets/RNAseq_report.Rmd   | 193 ++++++++++++++++++++++++++++++-------
 assets/references.bibtex   |  58 +++++++++++
 conf/test_smrnaseq.config  |  34 +++++++
 conf/test_smrnaseq2.config |  35 +++++++
 nextflow_schema.json       |   4 +-
 workflows/rnadeseq.nf      |   2 +-
 6 files changed, 287 insertions(+), 39 deletions(-)
 create mode 100644 conf/test_smrnaseq.config
 create mode 100644 conf/test_smrnaseq2.config

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 0fe78896..4c27ab9f 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -86,6 +86,35 @@ label_size <- function(dimension_int) {
     val <- max(c(2, 10-dimension_int/15))
     return(val)
 }
+
+# Function for creating a dataframe from smrnaseq files
+build_smrnaseq_df <- function(files) {
+    merged_mat <- F
+    for (f in files) {
+        file_tab <- read.table(f, sep="\t", row.names=1)
+        # Usually, the last row is weird and has the rowname *, remove it
+        if (grep("*", tail(rownames(file_tab), n=1), fixed=T)) {
+            file_tab <- head(file_tab, -1)
+        }
+
+        # If merged_mat already exists, add third hairpin column to it (second column after rownames)
+        if (is.data.frame(merged_mat)) {
+
+            # Check if rownames are the same
+            if (rownames(file_tab) != rownames(merged_mat)) {
+                stop(paste("Rownames of smrnaseq file", file_tab, "differ from the rownames of the other files; exiting..."))
+            }
+            merged_mat[[basename(f)]] <- file_tab[,2]
+        } else {
+
+            # Otherwise initiate it from third hairpin column and rownames
+            merged_mat <- data.frame(file_tab[,2])
+            rownames(merged_mat) <- rownames(file_tab)
+            colnames(merged_mat) <- c(basename(f))
+        }
+    }
+    merged_mat
+}
 ```
 
 ```{r param_validation, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
@@ -159,8 +188,8 @@ write_error_msg <- function(counts, qbicCodes) {
 }
 
 #Check necessary params
-if (!(params$input_type %in% c("featurecounts", "rsem", "salmon"))){
-    stop(paste0("Wrong input type ", params$input_type, ", must be one of [featurecounts, rsem, salmon]!"))
+if (!(params$input_type %in% c("featurecounts", "rsem", "salmon", "smrnaseq"))){
+    stop(paste0("Wrong input type ", params$input_type, ", must be one of [featurecounts, rsem, salmon, smrnaseq]!"))
 }
 if (params$input_type %in% c("rsem", "salmon") & !isProvided(params$path_gtf)){
     stop(paste0("For input type salmon, gtf file needs to be provided!\nIf using igenomes, please check that the entry for your genome contains a gtf file and otherwise provide one with `--gtf`."))
@@ -340,9 +369,60 @@ metadata$sampleName = paste(row.names(metadata),metadata$Secondary.Name,sep="_")
 row.names(metadata) = metadata$sampleName
 metadata_save <- metadata
 
+if (params$input_type == "smrnaseq") {
+
+    # Prepare files from smrnaseq, start with hairpins
+    hairpin_files <- file.path(gsub("/$", "", params$path_gene_counts), "hairpin", paste0(qbicCodes, "_mature_hairpin.sorted.bam.idxstats"))
+    if (!(all(file.exists(hairpin_files)))) {
+        missing_files <- "Could not find all of the specified <QBiC-code>_mature_hairpin.sorted.bam.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature_hairpin.sorted.bam.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
+        for (f in hairpin_files) {
+            if (!file.exists(f)) {
+                missing_files <- paste0(missing_files, f, ", ")
+            }
+        }
+        missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".")
+        stop(missing_files, call.=F)
+    }
+
+    # Non-hairpins next
+    other_files <- file.path(gsub("/$", "", params$path_gene_counts), "mature", paste0(qbicCodes, "_mature.sorted.bam.idxstats"))
+    if (!(all(file.exists(other_files)))) {
+        missing_files <- "Could not find all of the specified <QBiC-code>_mature.sorted.bam.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature.sorted.bam.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
+        for (f in other_files) {
+            if (!file.exists(f)) {
+                missing_files <- paste0(missing_files, f, ", ")
+            }
+        }
+        missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".")
+        stop(missing_files, call.=F)
+    }
+
+    # Merge hairpins and counts
+    hairpin_df <- build_smrnaseq_df(hairpin_files)
+    other_df <- build_smrnaseq_df(other_files)
+    colnames(other_df) <- colnames(hairpin_df)
+    count.table <- rbind(hairpin_df, other_df)
+
+
+    #TODO
+    # Need to order columns in count.table
+#    count.table <- count.table[, order(names(count.table))]
+
+
+    #count.table$Ensembl_ID <- rownames(count.table)
+    count.table <- cbind(Ensembl_ID=rownames(count.table), gene_name=rownames(count.table), count.table)
+    write.table(count.table, paste("differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double"))
+
+
+}
+
 # Load count table for FeatureCounts
 if (params$input_type == "featurecounts"){
     count.table <- read.table(params$path_gene_counts,  header = T,sep = "\t",na.strings =c("","NA"),quote=NULL,stringsAsFactors=F,dec=".",fill=TRUE,row.names=1)
+}
+
+# Processing is the same for FeatureCounts and the smrnaseq matrix that is built above
+if (params$input_type %in% c("featurecounts", "smrnaseq")) {
     count.table$Ensembl_ID <- row.names(count.table)
     drop <- c("Ensembl_ID","gene_name")
     gene_names <- count.table[,drop]
@@ -443,8 +523,10 @@ DT::datatable(df_QA_neat, options = list(scrollX  = "100%", scrollY = "400px", p
 
 ################## RUN DESEQ2 ######################################
 # Run DESeq function
-if (params$input_type == "featurecounts") {
+if (params$input_type %in% c("featurecounts", "smrnaseq")) {
+
     cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]]))))
+
 } else if (params$input_type %in% c("rsem", "salmon")) {
     ## Create a dataframe which consists of both the gene id and the transcript name
     gtf <- rtracklayer::import(params$path_gtf)
@@ -469,7 +551,7 @@ if (params$input_type == "featurecounts") {
         #-->the sample name is in the file name
         files <- file.path(gsub("/$", "", params$path_gene_counts), paste0(qbicCodes, ".genes.results"))
         if (!(all(file.exists(files)))) {
-            missing_files <- "DESeq2.R could not find all of the specified <QBiC-code>.genes.results files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>.genes.results file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
+            missing_files <- "Could not find all of the specified <QBiC-code>.genes.results files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>.genes.results file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
             for (f in files) {
                 if (!file.exists(f)) {
                     missing_files <- paste0(missing_files, f, ", ")
@@ -507,7 +589,7 @@ if (params$input_type == "featurecounts") {
         #-->the sample name is in the subfolder name, not in the file name!
         files <- file.path(gsub("/$", "", params$path_gene_counts), qbicCodes, "quant.sf")
         if (!(all(file.exists(files)))) {
-            missing_files <- "DESeq2.R could not find all of the specified <QBiC-code>/quant.sf files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>/quant.sf file is present in the gene_counts folder and <QBiC-code> subfolder and make sure that no typos exist. The following file(s) could not be found: "
+            missing_files <- "Could not find all of the specified <QBiC-code>/quant.sf files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>/quant.sf file is present in the gene_counts folder and <QBiC-code> subfolder and make sure that no typos exist. The following file(s) could not be found: "
             for (f in files) {
                 if (!file.exists(f)) {
                     missing_files <- paste0(missing_files, f, ", ")
@@ -533,7 +615,7 @@ if (params$input_type == "featurecounts") {
         cds <- DESeqDataSetFromTximport(txi=txi.salmon, colData =coldata, design = eval(parse(text=as.character(design[[1]]))))
     }
 } else {
-    stop(paste0("Invalid input type: ", params$input_type, "! Input type must be one of: featurecounts, rsem, salmon!"))
+    stop(paste0("Invalid input type: ", params$input_type, "! Input type must be one of: featurecounts, rsem, salmon, smrnaseq!"))
 }
 
 #Apply relevel if provided; this must be done before calling DESeq()
@@ -550,7 +632,7 @@ if (isProvided(params$path_relevel)) {
 cds <- DESeq(cds)
 
 # sizeFactors(cds) as indicator of library sequencing depth
-if (params$input_type == "featurecounts") {
+if (params$input_type %in% c("featurecounts", "smrnaseq")) {
     sf <- sizeFactors(cds)
 } else {
     # If tximport was used, sizeFactors doesn't work, so use workaround as described here: https://support.bioconductor.org/p/97676/
@@ -1171,7 +1253,7 @@ if (isProvided(params$path_contrast_matrix)){
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1231,7 +1313,7 @@ if (isProvided(params$path_contrast_list)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1273,7 +1355,7 @@ if (isProvided(params$path_contrast_pairs)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <-  DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
 
         # Round table before writing
@@ -1313,7 +1395,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <-  DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1695,7 +1777,7 @@ for (file in contrast_files){
     if (isProvided(params$set_background)) {
         gostres <- gost(query=q,
             organism=gost_id,
-            significant=TRUE,
+            significant=F,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -1711,7 +1793,7 @@ for (file in contrast_files){
         # Also run without bg for Venn diagrams
         gostres_nobg <- gost(query=q,
             organism=gost_id,
-            significant=TRUE,
+            significant=F,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -1725,7 +1807,7 @@ for (file in contrast_files){
     } else {
         gostres <- gost(query=q,
             organism=gost_id,
-            significant=TRUE,
+            significant=F,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -2000,9 +2082,12 @@ gost_text,
 The plot below summarizes the pathways that were found significantly enriched in DE genes for each contrast (padj value <= ", pval_text, ").
 Only contrasts for which an enriched pathway was found are shown. Both KEGG pathways and REACTOME (REAC) pathways are considered.
 Hover over the dots to reveal the pathway names. The table below provides more detail on all enriched pathways."))
+write(params$pathway_analysis, file="/sfs/9/ws/iivow01-test/rnadeseq/error/qw")
+
 ```
 
 ```{r gost, eval=params$pathway_analysis, out.height = "100%", out.width = "100%", warning=FALSE, message=FALSE}
+write(params$pathway_analysis, file="/sfs/9/ws/iivow01-test/rnadeseq/error/werwe")
 contrast_files <- list.files(path=paste0("differential_gene_expression/DE_genes_tables/"))
 q_list <- list()
 q_names <- c()
@@ -2030,7 +2115,7 @@ if (length(q_list) > 0) {
     if (isProvided(params$set_background)) {
         gostres <- gost(query=q_list,
                     organism=gost_id,
-                    significant=T,
+                    significant=F,
                     correction_method="fdr",
                     sources=datasources,
                     user_threshold=params$adj_pval_threshold,
@@ -2040,7 +2125,7 @@ if (length(q_list) > 0) {
     } else {
         gostres <- gost(query=q_list,
                     organism=gost_id,
-                    significant=T,
+                    significant=F,
                     correction_method="fdr",
                     sources=datasources,
                     user_threshold=params$adj_pval_threshold,
@@ -2074,6 +2159,8 @@ if (length(q_list) > 0) {
 } else {
     cat("No DE genes were found, pathway analysis not possible.")
     path_enrich=data.frame(list())
+    write("lahdladsh", file="/sfs/9/ws/iivow01-test/rnadeseq/error/eee")
+
 }
 ```
 
@@ -2109,6 +2196,7 @@ and the subfolder: `pathway_heatmaps`.
 # Summary and outlook
 
 ```{r quote, echo=FALSE, results='asis'}
+
 # Only add hyperlink to quote file if it was provided in the params
 quote_string <- ifelse(
     isProvided(params$path_quote),
@@ -2150,6 +2238,10 @@ if (software_versions_csv) {
         quant_tool <- "Salmon"
         quant_version <- as.character(version["SALMON_QUANT"][[1]]["salmon"])
         quant_cite <- "[@patro2017salmon]"
+    } else if (params$input_type == "smrnaseq") {
+        quant_tool <- "SAMtools"
+        quant_version <- as.character(version["SAMTOOLS_IDXSTATS"][[1]]["samtools"])
+        quant_cite <- "[@li2009sequence; @li2011statistical]"
     }
 }
 ```
@@ -2164,23 +2256,50 @@ cat("No software versions file was provided for primary analysis.")
 
 ```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
 
-# In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is obvious in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
-
+# In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is clearer in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
+# TODO remove F!
+if (params$input_type == "featurecounts" & F){
+    cat(paste0(
+    "## RNAseq data analysis
+
+    The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(software_versions_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
+    "` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
+    as.character(ifelse(software_versions_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
+    "` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
+    Subsequently, adapter trimming was conducted with `Trim Galore ",
+    as.character(ifelse(software_versions_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
+    "` [^4] [@krueger2012trim]. `STAR v",
+    as.character(ifelse(software_versions_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
+    "` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
+    The RNA-seq data quality control was performed with `RSeQC ",
+    as.character(ifelse(software_versions_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
+    "` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
+    quant_tool,
+    " ",
+    quant_version,
+    "` ",
+    quant_cite,
+    "."
+    ))
+} else if (params$input_type == "smrnaseq"){
 cat(paste0(
-"## RNAseq data analysis
-    
-The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(software_versions_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
-"` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
-as.character(ifelse(software_versions_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
-"` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
-Subsequently, adapter trimming was conducted with `Trim Galore ",
-as.character(ifelse(software_versions_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
-"` [^4] [@krueger2012trim]. `STAR v",
-as.character(ifelse(software_versions_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
-"` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
-The RNA-seq data quality control was performed with `RSeQC ",
-as.character(ifelse(software_versions_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
-"` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
+"## smRNA-seq data analysis
+
+The Nextflow-based nf-core pipeline `smrnaseq ", as.character(version["Workflow"][[1]]["nf-core/smrnaseq"]),
+"` [^6] was used for the smRNA-seq Bioinfomatics analysis. `FASTQC ",
+as.character(version["FASTQC"][[1]]["fastqc"]),
+"` [^3] [@andrews2010fastqc] was used to determine the quality of the miRNA samples. ",
+ifelse("TRIMGALORE" %in% names(version), paste0("Subsequently, adapter trimming was conducted with `Trim Galore ",
+as.character(version["TRIMGALORE"][[1]]["trimgalore"]),
+"` [^4] [@krueger2012trim]."),
+    ifelse("FASTP" %in% names(version), paste0("Subsequently, adapter trimming was conducted with `fastP ",
+    as.character(version["FASTP"][[1]]["fastp"]),
+    "` [^7] [@chen2018fastp; @chen2023ultrafast]."),
+        "")),
+" `Bowtie v", as.character(version["BOWTIE_MAP_HAIRPIN"][[1]]["bowtie"]), "` [@langmead2009ultrafast] aligner was used to map the reads that passed the quality control to the reference database.
+The smRNA-seq data quality control was performed with `miRTrace ",
+as.character(version["MIRTRACE_RUN"][[1]]["mirtrace"]),
+"` [^8] [@kang2018mirtrace] and read quantification of the features (e.g. genes) with `",
 quant_tool,
 " ",
 quant_version,
@@ -2188,15 +2307,14 @@ quant_version,
 quant_cite,
 "."
 ))
+}
+```
 
-```{r MQC_cite, echo=FALSE, results='asis', eval=software_versions_csv&isProvided(params$path_software_versions)}
-
+```{r MQC_cite, echo=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))&software_versions_csv&isProvided(params$path_software_versions)}
 cat(paste0("\nAn aggregation of the quality control for the RNAseq analysis was performed with `MultiQC ", as.character(version["MultiQC"]), "` [^2] [@ewels2016multiqc]."))
-
 ```
 
 ```{r software_versions_DE, echo=FALSE, results='asis'}
-
 cat(paste0(
 "## Differential gene expression analysis
 
@@ -2263,3 +2381,6 @@ sessioninfo::session_info()%>%
 [^4]: https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/
 [^5]: https://github.com/qbic-pipelines/rnadeseq
 
+[^6]: https://github.com/nf-core/smrnaseq
+[^7]: https://github.com/OpenGene/fastp
+[^8]: https://github.com/friedlanderlab/mirtrace
diff --git a/assets/references.bibtex b/assets/references.bibtex
index fdf26cc2..b16e7e87 100644
--- a/assets/references.bibtex
+++ b/assets/references.bibtex
@@ -216,3 +216,61 @@ journal="BMC Bioinformatics",
     year={2020},
     publisher={Nature Publishing Group}
 }
+@article{kang2018mirtrace,
+  title={miRTrace reveals the organismal origins of microRNA sequencing data},
+  author={Kang, Wenjing and Eldfjell, Yrin and Fromm, Bastian and Estivill, Xavier and Biryukova, Inna and Friedl{\"a}nder, Marc R},
+  journal={Genome biology},
+  volume={19},
+  number={1},
+  pages={1--15},
+  year={2018},
+  publisher={BioMed Central}
+}
+@article{langmead2009ultrafast,
+  title={Ultrafast and memory-efficient alignment of short DNA sequences to the human genome},
+  author={Langmead, Ben and Trapnell, Cole and Pop, Mihai and Salzberg, Steven L},
+  journal={Genome biology},
+  volume={10},
+  number={3},
+  pages={1--10},
+  year={2009},
+  publisher={BioMed Central}
+}
+@article{chen2018fastp,
+  title={fastp: an ultra-fast all-in-one FASTQ preprocessor},
+  author={Chen, Shifu and Zhou, Yanqing and Chen, Yaru and Gu, Jia},
+  journal={Bioinformatics},
+  volume={34},
+  number={17},
+  pages={i884--i890},
+  year={2018},
+  publisher={Oxford University Press}
+}
+@article{chen2023ultrafast,
+  title={Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp},
+  author={Chen, Shifu},
+  journal={iMeta},
+  pages={e107},
+  year={2023},
+  publisher={Wiley Online Library}
+}
+@article{li2009sequence,
+  title={The sequence alignment/map format and SAMtools},
+  author={Li, Heng and Handsaker, Bob and Wysoker, Alec and Fennell, Tim and Ruan, Jue and Homer, Nils and Marth, Gabor and Abecasis, Goncalo and Durbin, Richard and 1000 Genome Project Data Processing Subgroup},
+  journal={bioinformatics},
+  volume={25},
+  number={16},
+  pages={2078--2079},
+  year={2009},
+  publisher={Oxford University Press}
+}
+@article{li2011statistical,
+  title={A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data},
+  author={Li, Heng},
+  journal={Bioinformatics},
+  volume={27},
+  number={21},
+  pages={2987--2993},
+  year={2011},
+  publisher={Oxford University Press}
+}
diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config
new file mode 100644
index 00000000..c1851e8b
--- /dev/null
+++ b/conf/test_smrnaseq.config
@@ -0,0 +1,34 @@
+/*
+ * -------------------------------------------------
+ *  Nextflow config file for running tests
+ * -------------------------------------------------
+ * Defines bundled input files and everything required
+ * to run a fast and simple test. Use as follows:
+ *   nextflow run qbic-pipelines/rnadeseq -profile test_smrnaseq
+ */
+
+params {
+    config_profile_name = 'Test smrnaseq'
+    config_profile_description = 'Minimal test dataset to check pipeline function'
+    // Limit resources so that this can run on Travis
+    max_cpus = 2
+    max_memory = 6.GB
+    max_time = 48.h
+    // Input data
+    input = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/QBCOS_sample_preparations_smrnaseq_edit.tsv'
+    gene_counts = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_4/merged_gene_counts'
+    model = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/linear_model_timepoint.txt'
+    contrast_list = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/contrast_list_timepoint.tsv'
+    logFC_threshold = 1
+    //report_options = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/report_options.yml'
+    project_summary = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/QBCOS_20230612105025_summary.tsv'
+    software_versions = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_2/pipeline_info/software_versions.yml'
+    input_type = 'smrnaseq'
+    run_pathway_analysis = false
+    genome = 'GRCh37'
+    batch_effect = true
+    // species_library     = "org.Hs.eg.db"
+    // organism    = "hsapiens"
+    // keytype     = "ENSEMBL"
+    // gtf         = "s3://ngi-igenomes/igenomes/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
+}
diff --git a/conf/test_smrnaseq2.config b/conf/test_smrnaseq2.config
new file mode 100644
index 00000000..24c03a27
--- /dev/null
+++ b/conf/test_smrnaseq2.config
@@ -0,0 +1,35 @@
+/*
+ * -------------------------------------------------
+ *  Nextflow config file for running tests
+ * -------------------------------------------------
+ * Defines bundled input files and everything required
+ * to run a fast and simple test. Use as follows:
+ *   nextflow run qbic-pipelines/rnadeseq -profile test_smrnaseq
+ */
+
+params {
+    config_profile_name = 'Test smrnaseq'
+    config_profile_description = 'Minimal test dataset to check pipeline function'
+    // Limit resources so that this can run on Travis
+    max_cpus = 2
+    max_memory = 6.GB
+    max_time = 48.h
+    // Input data
+    input = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/QBCOS_sample_preparations_smrnaseq_edit.tsv'
+    gene_counts = '/sfs/9/ws/iivow01-test/rnadeseq/testdata/smrnaseq/merged_gene_counts_noHBLAK_introducedDE.txt'
+    model = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/linear_model_timepoint.txt'
+    contrast_list = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/contrast_list_timepoint.tsv'
+    logFC_threshold = 1
+    adj_pval_threshold = 0.05
+    //report_options = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/report_options.yml'
+    project_summary = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/QBCOS_20230612105025_summary.tsv'
+    software_versions = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_2/pipeline_info/software_versions.yml'
+    run_pathway_analysis = true
+    genome = 'GRCh37'
+    batch_effect = true
+    
+    // species_library     = "org.Hs.eg.db"
+    // organism    = "hsapiens"
+    // keytype     = "ENSEMBL"
+    // gtf         = "s3://ngi-igenomes/igenomes/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
+}
diff --git a/nextflow_schema.json b/nextflow_schema.json
index 3aec2e6a..c4eccfb4 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -13,11 +13,11 @@
                 "input_type": {
                     "type": "string",
                     "default": "featurecounts",
-                    "description": "Which type of input is provided, one of featurecounts, rsem, salmon"
+                    "description": "Which type of input is provided, one of featurecounts, rsem, salmon, smrnaseq"
                 },
                 "gene_counts": {
                     "type": "string",
-                    "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder."
+                    "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder. #TODO"
                 },
                 "input": {
                     "type": "string",
diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf
index f9b73a9c..ffbbde2c 100644
--- a/workflows/rnadeseq.nf
+++ b/workflows/rnadeseq.nf
@@ -15,7 +15,7 @@ def checkPathParamList = [
 for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } }
 
 // Check mandatory parameters
-if (!(params.input_type in ["featurecounts", "salmon", "rsem"])) { exit 1, 'Wrong input type ' + params.input_type + ', must be one of "featurecounts", "salmon", "rsem"!' }
+if (!(params.input_type in ["featurecounts", "salmon", "rsem", "smrnaseq"])) { exit 1, 'Wrong input type ' + params.input_type + ', must be one of "featurecounts", "salmon", "rsem", "smrnaseq"!' }
 
 ch_counts_path = Channel.fromPath(params.gene_counts)
 ch_metadata_file = Channel.fromPath(params.input)

From 8a64a90952ba3c1955d665495626641cf2ae875c Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 12:45:12 +0200
Subject: [PATCH 090/124] smrnaseq input works, updated changelog, test
 profiles and documentation

---
 .github/workflows/ci.yml   |  1 +
 CHANGELOG.md               |  1 +
 assets/RNAseq_report.Rmd   | 49 +++++++++++----------
 conf/test_smrnaseq.config  | 18 ++++----
 conf/test_smrnaseq2.config | 35 ---------------
 docs/output.md             | 13 +++---
 docs/usage.md              | 41 ++++++++++++++++--
 nextflow.config            |  1 +
 nextflow_schema.json       |  4 +-
 tests/test_smrnaseq.yml    | 89 ++++++++++++++++++++++++++++++++++++++
 tests/test_star_salmon.yml | 18 ++++----
 11 files changed, 180 insertions(+), 90 deletions(-)
 delete mode 100644 conf/test_smrnaseq2.config
 create mode 100644 tests/test_smrnaseq.yml

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index 6e833767..3996d01d 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -74,6 +74,7 @@ jobs:
             "test_contrast_list",
             "test_no_multiqc",
             "test_relevel",
+            "test_smrnaseq",
             "test_star_rsem",
             "test_star_salmon",
             "test_use_vst",
diff --git a/CHANGELOG.md b/CHANGELOG.md
index 2b5292b6..e07a79a6 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
+- [#213](https://github.com/qbic-pipelines/rnadeseq/pull/21) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
 - [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 4c27ab9f..4612c1e2 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -97,7 +97,7 @@ build_smrnaseq_df <- function(files) {
             file_tab <- head(file_tab, -1)
         }
 
-        # If merged_mat already exists, add third hairpin column to it (second column after rownames)
+        # If merged_mat already exists, add third matrix column to it (second column after rownames)
         if (is.data.frame(merged_mat)) {
 
             # Check if rownames are the same
@@ -107,7 +107,7 @@ build_smrnaseq_df <- function(files) {
             merged_mat[[basename(f)]] <- file_tab[,2]
         } else {
 
-            # Otherwise initiate it from third hairpin column and rownames
+            # Otherwise initiate it from third matrix column and rownames
             merged_mat <- data.frame(file_tab[,2])
             rownames(merged_mat) <- rownames(file_tab)
             colnames(merged_mat) <- c(basename(f))
@@ -372,9 +372,9 @@ metadata_save <- metadata
 if (params$input_type == "smrnaseq") {
 
     # Prepare files from smrnaseq, start with hairpins
-    hairpin_files <- file.path(gsub("/$", "", params$path_gene_counts), "hairpin", paste0(qbicCodes, "_mature_hairpin.sorted.bam.idxstats"))
+    hairpin_files <- file.path(gsub("/$", "", params$path_gene_counts), paste0(qbicCodes, "_mature_hairpin.sorted.idxstats"))
     if (!(all(file.exists(hairpin_files)))) {
-        missing_files <- "Could not find all of the specified <QBiC-code>_mature_hairpin.sorted.bam.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature_hairpin.sorted.bam.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
+        missing_files <- "Could not find all of the specified <QBiC-code>_mature_hairpin.sorted.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature_hairpin.sorted.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
         for (f in hairpin_files) {
             if (!file.exists(f)) {
                 missing_files <- paste0(missing_files, f, ", ")
@@ -385,9 +385,9 @@ if (params$input_type == "smrnaseq") {
     }
 
     # Non-hairpins next
-    other_files <- file.path(gsub("/$", "", params$path_gene_counts), "mature", paste0(qbicCodes, "_mature.sorted.bam.idxstats"))
+    other_files <- file.path(gsub("/$", "", params$path_gene_counts), paste0(qbicCodes, "_mature.sorted.idxstats"))
     if (!(all(file.exists(other_files)))) {
-        missing_files <- "Could not find all of the specified <QBiC-code>_mature.sorted.bam.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature.sorted.bam.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
+        missing_files <- "Could not find all of the specified <QBiC-code>_mature.sorted.idxstats files! Please check that for all QBiC codes in the metadata sheet, a <QBiC-code>_mature.sorted.idxstats file is present in the gene_counts folder and make sure that no typos exist. The following file(s) could not be found: "
         for (f in other_files) {
             if (!file.exists(f)) {
                 missing_files <- paste0(missing_files, f, ", ")
@@ -396,6 +396,9 @@ if (params$input_type == "smrnaseq") {
         missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".")
         stop(missing_files, call.=F)
     }
+    
+    # Combine files for later
+    files <- c(hairpin_files, other_files)
 
     # Merge hairpins and counts
     hairpin_df <- build_smrnaseq_df(hairpin_files)
@@ -403,13 +406,10 @@ if (params$input_type == "smrnaseq") {
     colnames(other_df) <- colnames(hairpin_df)
     count.table <- rbind(hairpin_df, other_df)
 
-
-    #TODO
     # Need to order columns in count.table
-#    count.table <- count.table[, order(names(count.table))]
-
+    count.table <- count.table[, order(names(count.table))]
 
-    #count.table$Ensembl_ID <- rownames(count.table)
+    count.table$Ensembl_ID <- rownames(count.table)
     count.table <- cbind(Ensembl_ID=rownames(count.table), gene_name=rownames(count.table), count.table)
     write.table(count.table, paste("differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double"))
 
@@ -441,8 +441,8 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) {
     count.table <- count.table[, order(names(count.table))]
 
     if (!identical(names(count.table),qbicCodes)) {
-    error_out <- write_error_msg(count.table, qbicCodes)
-    stop(paste0(error_out, "\n"), call.=F)
+        error_out <- write_error_msg(count.table, qbicCodes)
+        stop(paste0(error_out, "\n"), call.=F)
     }
     names(count.table) = metadata$sampleName
 
@@ -644,6 +644,11 @@ write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor
 # Write cds assay table to file
 write.table(counts(cds, normalized=T), paste0("differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv"), append=F, quote = F, sep = "\t", eol = "\n", na = "NA", dec = ".", row.names = T, col.names = T, qmethod = c("escape", "double"))
 
+# Or for smrnaseq save folder/file names to txt file
+if (params$input_type == "smrnaseq") {
+    write(files, file="differential_gene_expression/metadata/smrnaseq_files.txt")
+}
+
 # Or for rsem/salmon save folder/file names to txt file
 if (params$input_type == "rsem") {
     write(files, file="differential_gene_expression/metadata/star_rsem_files.txt")
@@ -1376,7 +1381,8 @@ if (isProvided(params$path_contrast_pairs)) {
 # Calculating DE genes for default contrasts (no contrast matrix or list or pairs provided)
 if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_list) & !isProvided(params$path_contrast_pairs)) {
     contrast_names <- coefficients[2:length(coefficients)]
-    contrast_names <- contrast_names[!startsWith(contrast_names, "batch_")]
+    contrast_names <- contrast_names[!startsWith(contrast_names, "batch")]
+
     for (contname in contrast_names) {
 
         results_DEseq_contrast <- results(cds, name=contname)
@@ -1777,7 +1783,7 @@ for (file in contrast_files){
     if (isProvided(params$set_background)) {
         gostres <- gost(query=q,
             organism=gost_id,
-            significant=F,
+            significant=T,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -1793,7 +1799,7 @@ for (file in contrast_files){
         # Also run without bg for Venn diagrams
         gostres_nobg <- gost(query=q,
             organism=gost_id,
-            significant=F,
+            significant=T,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -1807,7 +1813,7 @@ for (file in contrast_files){
     } else {
         gostres <- gost(query=q,
             organism=gost_id,
-            significant=F,
+            significant=T,
             correction_method="fdr",
             sources=datasources,
             evcodes=TRUE,
@@ -2082,12 +2088,9 @@ gost_text,
 The plot below summarizes the pathways that were found significantly enriched in DE genes for each contrast (padj value <= ", pval_text, ").
 Only contrasts for which an enriched pathway was found are shown. Both KEGG pathways and REACTOME (REAC) pathways are considered.
 Hover over the dots to reveal the pathway names. The table below provides more detail on all enriched pathways."))
-write(params$pathway_analysis, file="/sfs/9/ws/iivow01-test/rnadeseq/error/qw")
-
 ```
 
 ```{r gost, eval=params$pathway_analysis, out.height = "100%", out.width = "100%", warning=FALSE, message=FALSE}
-write(params$pathway_analysis, file="/sfs/9/ws/iivow01-test/rnadeseq/error/werwe")
 contrast_files <- list.files(path=paste0("differential_gene_expression/DE_genes_tables/"))
 q_list <- list()
 q_names <- c()
@@ -2115,7 +2118,7 @@ if (length(q_list) > 0) {
     if (isProvided(params$set_background)) {
         gostres <- gost(query=q_list,
                     organism=gost_id,
-                    significant=F,
+                    significant=T,
                     correction_method="fdr",
                     sources=datasources,
                     user_threshold=params$adj_pval_threshold,
@@ -2125,7 +2128,7 @@ if (length(q_list) > 0) {
     } else {
         gostres <- gost(query=q_list,
                     organism=gost_id,
-                    significant=F,
+                    significant=T,
                     correction_method="fdr",
                     sources=datasources,
                     user_threshold=params$adj_pval_threshold,
@@ -2159,8 +2162,6 @@ if (length(q_list) > 0) {
 } else {
     cat("No DE genes were found, pathway analysis not possible.")
     path_enrich=data.frame(list())
-    write("lahdladsh", file="/sfs/9/ws/iivow01-test/rnadeseq/error/eee")
-
 }
 ```
 
diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config
index c1851e8b..e421a1c3 100644
--- a/conf/test_smrnaseq.config
+++ b/conf/test_smrnaseq.config
@@ -15,20 +15,18 @@ params {
     max_memory = 6.GB
     max_time = 48.h
     // Input data
-    input = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/QBCOS_sample_preparations_smrnaseq_edit.tsv'
-    gene_counts = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_4/merged_gene_counts'
-    model = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/linear_model_timepoint.txt'
-    contrast_list = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/contrast_list_timepoint.tsv'
+    input = 'testdata/smrnaseq/samplesheet.tsv'
+    gene_counts = 'testdata/smrnaseq/counts'
+    model = '/home-link/iivow01/git/rnadeseq/testdata/smrnaseq/design.txt'
     logFC_threshold = 1
-    //report_options = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/report_options.yml'
-    project_summary = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/QBCOS_20230612105025_summary.tsv'
-    software_versions = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_2/pipeline_info/software_versions.yml'
+    project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
+    software_versions = '/home-link/iivow01/git/rnadeseq/testdata/smrnaseq/software_versions.yml'
+    genelist = 'testdata/smrnaseq/requested_genes.txt'
     input_type = 'smrnaseq'
-    run_pathway_analysis = false
+    run_pathway_analysis = true
     genome = 'GRCh37'
     batch_effect = true
+    set_background = false // This is solely to get some enriched pathways so that the github tests also check the pathway analysis output
     // species_library     = "org.Hs.eg.db"
     // organism    = "hsapiens"
-    // keytype     = "ENSEMBL"
-    // gtf         = "s3://ngi-igenomes/igenomes/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
 }
diff --git a/conf/test_smrnaseq2.config b/conf/test_smrnaseq2.config
deleted file mode 100644
index 24c03a27..00000000
--- a/conf/test_smrnaseq2.config
+++ /dev/null
@@ -1,35 +0,0 @@
-/*
- * -------------------------------------------------
- *  Nextflow config file for running tests
- * -------------------------------------------------
- * Defines bundled input files and everything required
- * to run a fast and simple test. Use as follows:
- *   nextflow run qbic-pipelines/rnadeseq -profile test_smrnaseq
- */
-
-params {
-    config_profile_name = 'Test smrnaseq'
-    config_profile_description = 'Minimal test dataset to check pipeline function'
-    // Limit resources so that this can run on Travis
-    max_cpus = 2
-    max_memory = 6.GB
-    max_time = 48.h
-    // Input data
-    input = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/QBCOS_sample_preparations_smrnaseq_edit.tsv'
-    gene_counts = '/sfs/9/ws/iivow01-test/rnadeseq/testdata/smrnaseq/merged_gene_counts_noHBLAK_introducedDE.txt'
-    model = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/linear_model_timepoint.txt'
-    contrast_list = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/deseq_input_1_batch_contrasts_all/contrast_list_timepoint.tsv'
-    logFC_threshold = 1
-    adj_pval_threshold = 0.05
-    //report_options = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/smrnaseq/report_options.yml'
-    project_summary = '/sfs/9/ws/qeajl01-QBCOS_smrna/data/QBCOS_20230612105025_summary.tsv'
-    software_versions = '/sfs/9/ws/qeajl01-QBCOS_smrna/results/smrnaseq/out_smrnaseq_2/pipeline_info/software_versions.yml'
-    run_pathway_analysis = true
-    genome = 'GRCh37'
-    batch_effect = true
-    
-    // species_library     = "org.Hs.eg.db"
-    // organism    = "hsapiens"
-    // keytype     = "ENSEMBL"
-    // gtf         = "s3://ngi-igenomes/igenomes/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf"
-}
diff --git a/docs/output.md b/docs/output.md
index bdc278d0..3c30fd18 100644
--- a/docs/output.md
+++ b/docs/output.md
@@ -27,8 +27,9 @@ This directory contains the zipped results. When unzipping them, the following s
   - `linear_model.txt`: linear model used for DE analysis.
   - `gene_list.txt`: provided list of interesting genes (if provided).
   - `star_[rsem/salmon]_files.txt`: list of counts files that were provided as input (only if `--input_type` is `rsem` or `salmon`).
+  - `smrnaseq_files.txt`: list of counts files that were provided as input (only if `--input_type` is `smrnaseq`).
 - `gene_counts_tables/`
-  - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline and used for the differential gene expression analysis (only if `--input_type` is `featurecounts`).
+  - `raw_gene_counts.txt`: raw gene counts table from the nf-core/rnaseq pipeline and used for the differential gene expression analysis (only if `--input_type` is `featurecounts` or `smrnaseq`; in the second case, will internally build a counts table from the input files, taking the third column of each .sorted.idxstats file (second column if counting rownames) and combining all values from both hairpin and non-hairpin mature files into one table).
   - `rlog_transformed_gene_counts.tsv`: normalized gene counts with the "regularized logarithm" approach. This normalization is used prior to PCA analysis and heatmap plotting of the gene counts (only when rlog transformation is used).
   - `vst_transformed_gene_counts.tsv`: normalized gene counts with the "variance stabilizing" transformation (only when vst transformation is used).
   - `sizeFactor_libraries.tsv`: size factors for each sample.
@@ -47,7 +48,7 @@ Pathway analysis with [gProfileR](https://biit.cs.ut.ee/gprofiler/gost) R packag
 
 **Output directory: `results/pathway_analysis/`**
 
-This directory contains the zipped pathway analysis results (`gProfileR.zip`). When unzipping them, a subfolder for each contrast used for the differential gene expression analysis is found. Inside each contrast folder, there is the following output:
+If pathway analysis was run, this directory contains the zipped pathway analysis results (`gProfileR.zip`). When unzipping them, a subfolder for each contrast used for the differential gene expression analysis is found. Inside each contrast folder, there is the following output:
 
 - `*_gost_pathway_venn_diagram.pdf/png`
   - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list.
@@ -66,8 +67,6 @@ QBiC report for RNAseq analysis.
 
 In this directory the zipped report is contained. This file is ready for upload to the project in the QBiC portal. Once unzipped, this directory contains:
 
-<!-- Here also remove the QC stuff? But then, nothing remains, right? -->
-
 - `Report.html`
   - QBiC report describing the RNAseq results.
 - `DESeq2/`
@@ -75,6 +74,6 @@ In this directory the zipped report is contained. This file is ready for upload
 - `QC/`
   - Quality control results. Contains:
   - `fastqc.zip`: zipped fastqc results for each sample.
-  - `multiqc_data`: data from the multiQC process of the RNAseq pipeline which is incorporated into the report.
-  - `multiqc_plots`: plots from the multiQC process of the RNAseq pipeline which are incorporated into the report.
-  - `qc_summary.tsv`: summary of the multiQC process in a table format.
+  - `multiqc_data`: If multiQC input was provided: data from the multiQC process of the RNAseq pipeline which is incorporated into the report.
+  - `multiqc_plots`: If multiQC input was provided: plots from the multiQC process of the RNAseq pipeline which are incorporated into the report.
+  - `qc_summary.tsv`: If multiQC input was provided: summary of the multiQC process in a table format.
diff --git a/docs/usage.md b/docs/usage.md
index 13587b1f..5801dfca 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -82,7 +82,7 @@ NXF_OPTS='-Xms1g -Xmx4g'
 
 ## Pre-requisites
 
-The `qbic-pipelines/rnadeseq` pipeline relies on the output from the `nf-core/rnaseq` pipeline. To be able to match the results of the `nf-core/rnaseq` pipeline with the metadata samplesheet containing the experimental design for the differential expression analysis, **the filenames of the fastq files used as input to the `qbic-pipelines/rnadeseq` pipeline, need to start with the corresponding QBiC codes!**. _E.g. QBICKXXXXX_original_file_name.fastq_. Once the filenames are corrected if necessary, you can run the `qbic-pipelines/rnadeseq` pipeline as usual.
+The `qbic-pipelines/rnadeseq` pipeline relies on the output from the `nf-core/rnaseq` pipeline or the `nf-core/smrnaseq` pipeline. To be able to match the results of the initial pipeline with the metadata samplesheet containing the experimental design for the differential expression analysis, **the filenames of the fastq files used as input to the `qbic-pipelines/rnadeseq` pipeline need to start with the corresponding QBiC codes!**. _E.g. QBICKXXXXX_original_file_name.fastq_. Once the filenames are corrected if necessary, you can run the `qbic-pipelines/rnadeseq` pipeline as usual.
 
 ## Running the pipeline
 
@@ -218,6 +218,41 @@ QDESQ083AC      Sample3 K562
 QDESQ084AK      Sample4 K562
 ```
 
+
+
+- OR a folder containing files with smrnaseq output (folder/sampleXXX_mature_hairpin.sorted.idxstats, folder/sampleXXX_mature_sorted.idxstats).
+  For smrnaseq, the --input_type parameter must be set to 'smrnaseq'.
+  For example:
+
+```bash
+--gene_counts 'path/to/smrnaseq_folder' --metadata 'path/to/smrnaseq_metadata.tsv'
+
+tree testdata/smrnaseq/counts/
+testdata/smrnaseq/counts/
+├── Clone1_NN1_mature.sorted.idxstats
+├── Clone1_NN1_mature_hairpin.sorted.idxstats
+├── Clone1_NN3_mature.sorted.idxstats
+├── Clone1_NN3_mature_hairpin.sorted.idxstats
+├── Clone9_NN1_mature.sorted.idxstats
+├── Clone9_NN1_mature_hairpin.sorted.idxstats
+.
+.
+.
+
+head 'path/to/smrnaseq_metadata.tsv'
+```
+
+```tsv
+QBiC Code       Secondary Name  Condition: cellline
+QDESQ081AU      Sample1 GM12878
+QDESQ082A4      Sample2 GM12878
+QDESQ083AC      Sample3 K562
+QDESQ084AK      Sample4 K562
+```
+
+
+
+
 ### `--input`
 
 Metadata table/samplesheet (TSV) is the "Sample_preparations_sheet.tsv" that can be directly downloaded from the qPortal --> Browser. Rows are samples and columns contain sample grouping. Important columns are:
@@ -344,7 +379,7 @@ Set this flag to run pathway analysis, otherwise, this step will be skipped.
 
 ### `--input_type`
 
-This tells the pipeline which type of input dataset is provided. Must be one of 'featurecounts', 'rsem', 'salmon', default: featurecounts.
+This tells the pipeline which type of input dataset is provided. Must be one of 'featurecounts', 'rsem', 'salmon', 'smrnaseq', default: featurecounts.
 
 ### `--multiqc`
 
@@ -393,7 +428,7 @@ Which genome to use for analysis, e.g. GRCh37; see `/conf/igenomes.config` for w
 
 ### `--gtf`
 
-GTF file to be used for DESeq if input is rsem or salmon, not necessary for featurecounts.
+GTF file to be used for DESeq if input is rsem or salmon, not necessary for featurecounts or smrnaseq.
 
 ### `--organism`
 
diff --git a/nextflow.config b/nextflow.config
index fd1595d6..866a45e4 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -198,6 +198,7 @@ profiles {
     test_contrast_list { includeConfig 'conf/test_contrast_list.config' }
     test_relevel { includeConfig 'conf/test_relevel.config' }
     test_use_vst { includeConfig 'conf/test_use_vst.config' }
+    test_smrnaseq { includeConfig 'conf/test_smrnaseq.config' }
     test_star_salmon { includeConfig 'conf/test_star_salmon.config' }
     test_star_rsem { includeConfig 'conf/test_star_rsem.config' }
     test_batcheffect { includeConfig 'conf/test_batcheffect.config' }
diff --git a/nextflow_schema.json b/nextflow_schema.json
index c4eccfb4..e992d9cd 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -13,11 +13,11 @@
                 "input_type": {
                     "type": "string",
                     "default": "featurecounts",
-                    "description": "Which type of input is provided, one of featurecounts, rsem, salmon, smrnaseq"
+                    "description": "Which type of input is provided, one of featurecounts, rsem, salmon, smrnaseq."
                 },
                 "gene_counts": {
                     "type": "string",
-                    "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder. #TODO"
+                    "description": "If input_type = featurecounts: Raw count table (TSV). Columns are samples and rows are genes. 1st column Ensembl_ID, 2nd column gene_name. If input_type = rsem or input_type = salmon: Path to rsem/salmon output folder. If input_type = smrnaseq: Path to smrnaseq output folder containing hairpin and non-hairpin mature .sorted.idxstats files."
                 },
                 "input": {
                     "type": "string",
diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml
new file mode 100644
index 00000000..493b7b36
--- /dev/null
+++ b/tests/test_smrnaseq.yml
@@ -0,0 +1,89 @@
+- name: Run test profile with smrnaseq
+  command: nextflow run main.nf -profile test_smrnaseq,docker --citest true --round_DE 3
+  tags:
+    - test_star_salmon
+  files:
+    - path: results_test/differential_gene_expression/metadata/smrnaseq_files.txt
+      md5sum: ce3cc117fc953a8ff4d17d0118123faf
+    - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_clone9_vs_clone1.tsv
+      md5sum: 46ca6c2ec1046e088853c96b28cd30e3
+    - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_control_vs_clone1.tsv
+      md5sum: 74214d33037aeded1bd0c839973a9e4c
+    - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
+      md5sum: 9f4950203e56a94235b00a1d554fa78c
+    - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv
+    - path: results_test/differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-LET-7C_hsa-let-7c.pdf
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-LET-7C_hsa-let-7c.png
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-LET-7C_hsa-let-7c.svg
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-100_hsa-mir-100.pdf
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-100_hsa-mir-100.png
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-100_hsa-mir-100.svg
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-125B-1_hsa-mir-125b-1.pdf
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-125B-1_hsa-mir-125b-1.png
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-125B-1_hsa-mir-125b-1.svg
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-200A_hsa-mir-200a.pdf
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-200A_hsa-mir-200a.png
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-200A_hsa-mir-200a.svg
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-505_hsa-mir-505.pdf
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-505_hsa-mir-505.png
+    - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/HSA-MIR-505_hsa-mir-505.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_clone9_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_clone9_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_clone9_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_control_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_control_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_treatment_control_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_clone9_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_clone9_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_clone9_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_control_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_control_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_treatment_control_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_clone9_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_clone9_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_clone9_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_control_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_control_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/number.of.rejections_condition_treatment_control_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_clone9_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_clone9_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_clone9_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_control_vs_clone1.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_control_vs_clone1.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/dependency_small.pval_mean_normal.counts_condition_treatment_control_vs_clone1.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Dispersion_plot.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Dispersion_plot.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Dispersion_plot.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/Effects_of_transformations_on_the_variance.svg
+    - path: results_test/differential_gene_expression/plots/Heatmaps_of_distances.pdf
+    - path: results_test/differential_gene_expression/plots/Heatmaps_of_distances.png
+    - path: results_test/differential_gene_expression/plots/Heatmaps_of_distances.svg
+    - path: results_test/differential_gene_expression/plots/PCA_plot.pdf
+    - path: results_test/differential_gene_expression/plots/PCA_plot.png
+    - path: results_test/differential_gene_expression/plots/PCA_plot.svg
+    - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.pdf
+    - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.png
+    - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.svg
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.pdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.png
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.svg
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_results.tsv
+      md5sum: b5c4c68a304f08a54b2b9d8969334e8c
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_pathway_enrichment_results.tsv
+      md5sum: ab088ff01a4e3991be8a424e676afcdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.pdf
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.png
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.svg
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_results.tsv
+      md5sum: 4a2ed390ae3e9fea628bd9c014e88cc6
+    - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_pathway_enrichment_results.tsv
+      md5sum: db2149ba5f2dbb6d4aed3f6284072f39
+    - path: results_test/RNAseq_report.html
diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml
index 20b498cc..dbe49c9a 100644
--- a/tests/test_star_salmon.yml
+++ b/tests/test_star_salmon.yml
@@ -19,15 +19,15 @@
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL10_ENSG00000136634.pdf
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL10_ENSG00000136634.png
     - path: results_test/differential_gene_expression/plots/boxplots_requested_genes/IL10_ENSG00000136634.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_H1_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_H1_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_H1_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_K562_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_K562_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_K562_vs_GM12878.svg
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.pdf
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.png
-    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots//all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_H1_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_H1_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_H1_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_K562_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_K562_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_K562_vs_GM12878.svg
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.pdf
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.png
+    - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_condition_cellline_MCF7_vs_GM12878.svg
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.pdf
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.png
     - path: results_test/differential_gene_expression/plots/further_diagnostics_plots/histogram_of_p.values_condition_cellline_H1_vs_GM12878.svg

From 6c914e400a5055d4eeb455494c3408f23dd2bb32 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 12:49:39 +0200
Subject: [PATCH 091/124] prettier

---
 .prettierignore | 1 +
 docs/usage.md   | 5 -----
 2 files changed, 1 insertion(+), 5 deletions(-)

diff --git a/.prettierignore b/.prettierignore
index 437d763d..88a6ffb4 100644
--- a/.prettierignore
+++ b/.prettierignore
@@ -6,6 +6,7 @@ work/
 data/
 results/
 .DS_Store
+testdata/
 testing/
 testing*
 *.pyc
diff --git a/docs/usage.md b/docs/usage.md
index 5801dfca..2f6719e4 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -218,8 +218,6 @@ QDESQ083AC      Sample3 K562
 QDESQ084AK      Sample4 K562
 ```
 
-
-
 - OR a folder containing files with smrnaseq output (folder/sampleXXX_mature_hairpin.sorted.idxstats, folder/sampleXXX_mature_sorted.idxstats).
   For smrnaseq, the --input_type parameter must be set to 'smrnaseq'.
   For example:
@@ -250,9 +248,6 @@ QDESQ083AC      Sample3 K562
 QDESQ084AK      Sample4 K562
 ```
 
-
-
-
 ### `--input`
 
 Metadata table/samplesheet (TSV) is the "Sample_preparations_sheet.tsv" that can be directly downloaded from the qPortal --> Browser. Rows are samples and columns contain sample grouping. Important columns are:

From daaf691a470c41f4dfc12966c80a759f95562dc6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 12:50:05 +0200
Subject: [PATCH 092/124] added smrnaseq testdata

---
 .../counts/Clone1_NN1_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Clone1_NN1_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Clone1_NN3_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Clone1_NN3_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Clone9_NN1_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Clone9_NN1_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Clone9_NN2_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Clone9_NN2_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Clone9_NN3_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Clone9_NN3_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Control_N1_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Control_N1_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Control_N2_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Control_N2_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 .../counts/Control_N3_mature.sorted.idxstats  | 2589 +++++++++++++++++
 .../Control_N3_mature_hairpin.sorted.idxstats | 1882 ++++++++++++
 testdata/smrnaseq/design.txt                  |    1 +
 testdata/smrnaseq/requested_genes.txt         |    5 +
 testdata/smrnaseq/samplesheet.tsv             |    9 +
 testdata/smrnaseq/software_versions.yml       |   64 +
 20 files changed, 35847 insertions(+)
 create mode 100644 testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats
 create mode 100644 testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats
 create mode 100644 testdata/smrnaseq/design.txt
 create mode 100644 testdata/smrnaseq/requested_genes.txt
 create mode 100644 testdata/smrnaseq/samplesheet.tsv
 create mode 100644 testdata/smrnaseq/software_versions.yml

diff --git a/testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats b/testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats
new file mode 100644
index 00000000..8ccd1462
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
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+hsa-miR-8089	24	0	0
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+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	2193613
diff --git a/testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..5e20365d
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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diff --git a/testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats b/testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats
new file mode 100644
index 00000000..095d2a32
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
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+hsa-miR-6830-3p	23	0	0
+hsa-miR-6831-5p	24	0	0
+hsa-miR-6831-3p	21	0	0
+hsa-miR-6832-5p	23	0	0
+hsa-miR-6832-3p	20	0	0
+hsa-miR-6833-5p	22	0	0
+hsa-miR-6833-3p	21	0	0
+hsa-miR-6834-5p	21	0	0
+hsa-miR-6834-3p	20	0	0
+hsa-miR-6835-5p	22	0	0
+hsa-miR-6835-3p	23	0	0
+hsa-miR-6780b-5p	23	0	0
+hsa-miR-6780b-3p	21	0	0
+hsa-miR-6836-5p	22	0	0
+hsa-miR-6836-3p	21	0	0
+hsa-miR-6837-5p	21	0	0
+hsa-miR-6837-3p	23	0	0
+hsa-miR-6838-5p	22	0	0
+hsa-miR-6838-3p	20	0	0
+hsa-miR-6839-5p	22	0	0
+hsa-miR-6839-3p	22	0	0
+hsa-miR-6840-5p	24	0	0
+hsa-miR-6840-3p	22	0	0
+hsa-miR-6841-5p	22	0	0
+hsa-miR-6841-3p	21	0	0
+hsa-miR-6842-5p	22	0	0
+hsa-miR-6842-3p	22	0	0
+hsa-miR-6843-3p	21	0	0
+hsa-miR-6844	21	0	0
+hsa-miR-6845-5p	19	0	0
+hsa-miR-6845-3p	21	0	0
+hsa-miR-6846-5p	22	0	0
+hsa-miR-6846-3p	21	0	0
+hsa-miR-6847-5p	23	0	0
+hsa-miR-6847-3p	22	0	0
+hsa-miR-6848-5p	23	0	0
+hsa-miR-6848-3p	19	0	0
+hsa-miR-6849-5p	23	0	0
+hsa-miR-6849-3p	22	0	0
+hsa-miR-6850-5p	22	0	0
+hsa-miR-6850-3p	20	0	0
+hsa-miR-6851-5p	23	0	0
+hsa-miR-6851-3p	21	0	0
+hsa-miR-6852-5p	21	0	0
+hsa-miR-6852-3p	17	0	0
+hsa-miR-6853-5p	24	0	0
+hsa-miR-6853-3p	22	0	0
+hsa-miR-6854-5p	24	0	0
+hsa-miR-6854-3p	21	0	0
+hsa-miR-6855-5p	25	0	0
+hsa-miR-6855-3p	22	0	0
+hsa-miR-6856-5p	24	0	0
+hsa-miR-6856-3p	22	0	0
+hsa-miR-6857-5p	22	0	0
+hsa-miR-6857-3p	21	0	0
+hsa-miR-6858-5p	22	0	0
+hsa-miR-6858-3p	22	0	0
+hsa-miR-6859-5p	23	0	0
+hsa-miR-6859-3p	23	0	0
+hsa-miR-6769b-5p	23	0	0
+hsa-miR-6769b-3p	21	0	0
+hsa-miR-6860	22	0	0
+hsa-miR-6861-5p	22	0	0
+hsa-miR-6861-3p	18	0	0
+hsa-miR-6862-5p	22	0	0
+hsa-miR-6862-3p	24	0	0
+hsa-miR-6863	23	0	0
+hsa-miR-6864-5p	24	0	0
+hsa-miR-6864-3p	21	0	0
+hsa-miR-6865-5p	23	0	0
+hsa-miR-6865-3p	21	0	0
+hsa-miR-6866-5p	23	0	0
+hsa-miR-6866-3p	22	0	0
+hsa-miR-6867-5p	23	0	0
+hsa-miR-6867-3p	21	0	0
+hsa-miR-6868-5p	22	0	0
+hsa-miR-6868-3p	21	0	0
+hsa-miR-6869-5p	22	0	0
+hsa-miR-6869-3p	21	0	0
+hsa-miR-6870-5p	19	0	0
+hsa-miR-6870-3p	22	0	0
+hsa-miR-6871-5p	22	0	0
+hsa-miR-6871-3p	22	0	0
+hsa-miR-6872-5p	20	0	0
+hsa-miR-6872-3p	21	0	0
+hsa-miR-6873-5p	22	0	0
+hsa-miR-6873-3p	23	0	0
+hsa-miR-6874-5p	23	0	0
+hsa-miR-6874-3p	23	0	0
+hsa-miR-6875-5p	21	0	0
+hsa-miR-6875-3p	22	0	0
+hsa-miR-6876-5p	22	0	0
+hsa-miR-6876-3p	24	0	0
+hsa-miR-6877-5p	22	0	0
+hsa-miR-6877-3p	21	0	0
+hsa-miR-6878-5p	23	0	0
+hsa-miR-6878-3p	21	0	0
+hsa-miR-6879-5p	22	0	0
+hsa-miR-6879-3p	21	0	0
+hsa-miR-6880-5p	22	0	0
+hsa-miR-6880-3p	21	0	0
+hsa-miR-6881-5p	22	0	0
+hsa-miR-6881-3p	22	0	0
+hsa-miR-6882-5p	22	0	0
+hsa-miR-6882-3p	24	0	0
+hsa-miR-6883-5p	22	0	0
+hsa-miR-6883-3p	22	0	0
+hsa-miR-6884-5p	22	0	0
+hsa-miR-6884-3p	23	0	0
+hsa-miR-6885-5p	25	0	0
+hsa-miR-6885-3p	21	0	0
+hsa-miR-6886-5p	21	0	0
+hsa-miR-6886-3p	21	0	0
+hsa-miR-6887-5p	23	0	0
+hsa-miR-6887-3p	21	0	0
+hsa-miR-6888-5p	20	0	0
+hsa-miR-6888-3p	21	0	0
+hsa-miR-6889-5p	23	0	0
+hsa-miR-6889-3p	20	0	0
+hsa-miR-6890-5p	21	0	0
+hsa-miR-6890-3p	20	0	0
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+hsa-miR-8485	21	0	0
+hsa-miR-9500	19	0	0
+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	1649307
diff --git a/testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..399b6528
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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+*	0	0	1641154
diff --git a/testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats
new file mode 100644
index 00000000..022f65f0
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
+hsa-let-7a-5p	22	1	0
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+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	2495471
diff --git a/testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..f296b34f
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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diff --git a/testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats
new file mode 100644
index 00000000..71022913
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
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+hsa-miR-6846-5p	22	0	0
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+hsa-miR-6847-5p	23	0	0
+hsa-miR-6847-3p	22	0	0
+hsa-miR-6848-5p	23	0	0
+hsa-miR-6848-3p	19	0	0
+hsa-miR-6849-5p	23	0	0
+hsa-miR-6849-3p	22	0	0
+hsa-miR-6850-5p	22	0	0
+hsa-miR-6850-3p	20	0	0
+hsa-miR-6851-5p	23	0	0
+hsa-miR-6851-3p	21	0	0
+hsa-miR-6852-5p	21	0	0
+hsa-miR-6852-3p	17	0	0
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+hsa-miR-6853-3p	22	0	0
+hsa-miR-6854-5p	24	0	0
+hsa-miR-6854-3p	21	0	0
+hsa-miR-6855-5p	25	0	0
+hsa-miR-6855-3p	22	0	0
+hsa-miR-6856-5p	24	0	0
+hsa-miR-6856-3p	22	0	0
+hsa-miR-6857-5p	22	0	0
+hsa-miR-6857-3p	21	0	0
+hsa-miR-6858-5p	22	0	0
+hsa-miR-6858-3p	22	0	0
+hsa-miR-6859-5p	23	0	0
+hsa-miR-6859-3p	23	0	0
+hsa-miR-6769b-5p	23	0	0
+hsa-miR-6769b-3p	21	0	0
+hsa-miR-6860	22	0	0
+hsa-miR-6861-5p	22	0	0
+hsa-miR-6861-3p	18	0	0
+hsa-miR-6862-5p	22	0	0
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+hsa-miR-6863	23	0	0
+hsa-miR-6864-5p	24	0	0
+hsa-miR-6864-3p	21	0	0
+hsa-miR-6865-5p	23	0	0
+hsa-miR-6865-3p	21	0	0
+hsa-miR-6866-5p	23	0	0
+hsa-miR-6866-3p	22	0	0
+hsa-miR-6867-5p	23	0	0
+hsa-miR-6867-3p	21	0	0
+hsa-miR-6868-5p	22	0	0
+hsa-miR-6868-3p	21	0	0
+hsa-miR-6869-5p	22	0	0
+hsa-miR-6869-3p	21	0	0
+hsa-miR-6870-5p	19	0	0
+hsa-miR-6870-3p	22	0	0
+hsa-miR-6871-5p	22	0	0
+hsa-miR-6871-3p	22	0	0
+hsa-miR-6872-5p	20	0	0
+hsa-miR-6872-3p	21	0	0
+hsa-miR-6873-5p	22	0	0
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+hsa-miR-6874-5p	23	0	0
+hsa-miR-6874-3p	23	0	0
+hsa-miR-6875-5p	21	0	0
+hsa-miR-6875-3p	22	0	0
+hsa-miR-6876-5p	22	0	0
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+hsa-miR-6877-5p	22	0	0
+hsa-miR-6877-3p	21	0	0
+hsa-miR-6878-5p	23	0	0
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+hsa-miR-6879-5p	22	0	0
+hsa-miR-6879-3p	21	0	0
+hsa-miR-6880-5p	22	0	0
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+hsa-miR-6881-5p	22	0	0
+hsa-miR-6881-3p	22	0	0
+hsa-miR-6882-5p	22	0	0
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+hsa-miR-6885-5p	25	0	0
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+hsa-miR-6887-5p	23	0	0
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+hsa-miR-6888-5p	20	0	0
+hsa-miR-6888-3p	21	0	0
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+hsa-miR-6890-5p	21	0	0
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+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	1922831
diff --git a/testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..7a7ce005
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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+*	0	0	1909506
diff --git a/testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats
new file mode 100644
index 00000000..3d15982c
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
+hsa-let-7a-5p	22	0	0
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+*	0	0	1487807
diff --git a/testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..37948f60
--- /dev/null
+++ b/testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
+hsa-let-7a-1	80	269	0
+hsa-let-7a-2	72	357	0
+hsa-let-7a-3	74	16	0
+hsa-let-7b	83	243	0
+hsa-let-7c	84	326	0
+hsa-let-7d	87	12	0
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diff --git a/testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats b/testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats
new file mode 100644
index 00000000..c6c73776
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
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+hsa-miR-6859-3p	23	0	0
+hsa-miR-6769b-5p	23	0	0
+hsa-miR-6769b-3p	21	0	0
+hsa-miR-6860	22	0	0
+hsa-miR-6861-5p	22	0	0
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+hsa-miR-6864-5p	24	0	0
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+hsa-miR-6865-3p	21	0	0
+hsa-miR-6866-5p	23	0	0
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+hsa-miR-6867-5p	23	0	0
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+hsa-miR-6868-3p	21	0	0
+hsa-miR-6869-5p	22	0	0
+hsa-miR-6869-3p	21	0	0
+hsa-miR-6870-5p	19	0	0
+hsa-miR-6870-3p	22	0	0
+hsa-miR-6871-5p	22	0	0
+hsa-miR-6871-3p	22	0	0
+hsa-miR-6872-5p	20	0	0
+hsa-miR-6872-3p	21	0	0
+hsa-miR-6873-5p	22	0	0
+hsa-miR-6873-3p	23	0	0
+hsa-miR-6874-5p	23	0	0
+hsa-miR-6874-3p	23	0	0
+hsa-miR-6875-5p	21	0	0
+hsa-miR-6875-3p	22	0	0
+hsa-miR-6876-5p	22	0	0
+hsa-miR-6876-3p	24	0	0
+hsa-miR-6877-5p	22	0	0
+hsa-miR-6877-3p	21	0	0
+hsa-miR-6878-5p	23	0	0
+hsa-miR-6878-3p	21	0	0
+hsa-miR-6879-5p	22	0	0
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+hsa-miR-6880-5p	22	0	0
+hsa-miR-6880-3p	21	0	0
+hsa-miR-6881-5p	22	0	0
+hsa-miR-6881-3p	22	0	0
+hsa-miR-6882-5p	22	0	0
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+hsa-miR-6883-3p	22	0	0
+hsa-miR-6884-5p	22	0	0
+hsa-miR-6884-3p	23	0	0
+hsa-miR-6885-5p	25	0	0
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+hsa-miR-6886-5p	21	0	0
+hsa-miR-6886-3p	21	0	0
+hsa-miR-6887-5p	23	0	0
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+hsa-miR-6888-5p	20	0	0
+hsa-miR-6888-3p	21	0	0
+hsa-miR-6889-5p	23	0	0
+hsa-miR-6889-3p	20	0	0
+hsa-miR-6890-5p	21	0	0
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+hsa-miR-8485	21	0	0
+hsa-miR-9500	19	0	0
+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	1713517
diff --git a/testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..3d601dcd
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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+hsa-mir-7641-2	53	337	0
+hsa-mir-7702	59	0	0
+hsa-mir-7703	77	0	0
+hsa-mir-7704	59	0	0
+hsa-mir-7705	57	0	0
+hsa-mir-7706	67	0	0
+hsa-mir-7843	79	0	0
+hsa-mir-4433b	102	0	0
+hsa-mir-1273h	116	0	0
+hsa-mir-6516	81	0	0
+hsa-mir-7844	122	0	0
+hsa-mir-7845	99	0	0
+hsa-mir-7846	96	0	0
+hsa-mir-7847	103	0	0
+hsa-mir-7848	101	0	0
+hsa-mir-7849	106	0	0
+hsa-mir-7850	79	0	0
+hsa-mir-7851	160	0	0
+hsa-mir-7852	82	0	0
+hsa-mir-7853	132	0	0
+hsa-mir-7854	65	0	0
+hsa-mir-7855	61	0	0
+hsa-mir-7856	56	0	0
+hsa-mir-548ba	56	0	0
+hsa-mir-7973-1	76	0	0
+hsa-mir-7973-2	76	0	0
+hsa-mir-7974	79	0	0
+hsa-mir-7975	68	0	0
+hsa-mir-7976	66	0	0
+hsa-mir-7977	49	0	0
+hsa-mir-7978	59	0	0
+hsa-mir-8052	69	0	0
+hsa-mir-8053	75	0	0
+hsa-mir-8054	86	0	0
+hsa-mir-8055	97	0	0
+hsa-mir-8056	82	0	0
+hsa-mir-8057	69	0	0
+hsa-mir-8058	89	0	0
+hsa-mir-8059	81	0	0
+hsa-mir-8060	76	0	0
+hsa-mir-8061	75	0	0
+hsa-mir-8062	85	0	0
+hsa-mir-8063	81	0	0
+hsa-mir-8064	90	0	0
+hsa-mir-8065	100	0	0
+hsa-mir-8066	78	0	0
+hsa-mir-8067	77	0	0
+hsa-mir-8068	68	0	0
+hsa-mir-8069-1	86	0	0
+hsa-mir-8070	88	0	0
+hsa-mir-8071-1	65	0	0
+hsa-mir-8072	80	0	0
+hsa-mir-8073	72	0	0
+hsa-mir-8074	81	0	0
+hsa-mir-8075	80	0	0
+hsa-mir-8076	83	0	0
+hsa-mir-8077	75	0	0
+hsa-mir-8078	84	0	0
+hsa-mir-8079	72	0	0
+hsa-mir-8080	89	0	0
+hsa-mir-8081	95	0	0
+hsa-mir-8082	81	0	0
+hsa-mir-8083	89	0	0
+hsa-mir-8084	89	0	0
+hsa-mir-8085	65	0	0
+hsa-mir-8086	93	5	0
+hsa-mir-8087	78	0	0
+hsa-mir-8088	86	0	0
+hsa-mir-8089	82	0	0
+hsa-mir-6862-2	70	0	0
+hsa-mir-8071-2	65	0	0
+hsa-mir-6770-2	60	0	0
+hsa-mir-6770-3	60	0	0
+hsa-mir-6859-2	68	0	0
+hsa-mir-6859-3	68	0	0
+hsa-mir-8485	91	0	0
+hsa-mir-9500	65	0	0
+hsa-mir-548bb	66	0	0
+hsa-mir-3179-4	84	0	0
+hsa-mir-1244-4	85	0	0
+hsa-mir-3648-2	180	0	0
+hsa-mir-3670-3	65	0	0
+hsa-mir-3670-4	65	0	0
+hsa-mir-3687-2	61	0	0
+hsa-mir-6724-2	92	0	0
+hsa-mir-6724-3	92	0	0
+hsa-mir-6724-4	92	0	0
+hsa-mir-8069-2	86	0	0
+hsa-mir-941-5	72	0	0
+hsa-mir-6859-4	68	0	0
+hsa-mir-5701-3	82	0	0
+*	0	0	1681660
diff --git a/testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats b/testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats
new file mode 100644
index 00000000..e29756fd
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
+hsa-let-7a-5p	22	0	0
+hsa-let-7a-3p	21	0	0
+hsa-let-7a-2-3p	22	0	0
+hsa-let-7b-5p	22	0	0
+hsa-let-7b-3p	22	0	0
+hsa-let-7c-5p	22	0	0
+hsa-let-7c-3p	22	0	0
+hsa-let-7d-5p	22	0	0
+hsa-let-7d-3p	22	0	0
+hsa-let-7e-5p	22	0	0
+hsa-let-7e-3p	22	0	0
+hsa-let-7f-5p	22	0	0
+hsa-let-7f-1-3p	22	0	0
+hsa-let-7f-2-3p	22	0	0
+hsa-miR-15a-5p	22	0	0
+hsa-miR-15a-3p	22	0	0
+hsa-miR-16-5p	22	0	0
+hsa-miR-16-1-3p	22	0	0
+hsa-miR-17-5p	23	0	0
+hsa-miR-17-3p	22	0	0
+hsa-miR-18a-5p	23	0	0
+hsa-miR-18a-3p	23	0	0
+hsa-miR-19a-5p	22	0	0
+hsa-miR-19a-3p	23	0	0
+hsa-miR-19b-1-5p	23	0	0
+hsa-miR-19b-3p	23	0	0
+hsa-miR-19b-2-5p	22	0	0
+hsa-miR-20a-5p	23	0	0
+hsa-miR-20a-3p	22	0	0
+hsa-miR-21-5p	22	0	0
+hsa-miR-21-3p	21	0	0
+hsa-miR-22-5p	22	0	0
+hsa-miR-22-3p	22	0	0
+hsa-miR-23a-5p	22	0	0
+hsa-miR-23a-3p	21	0	0
+hsa-miR-24-1-5p	22	0	0
+hsa-miR-24-3p	22	0	0
+hsa-miR-24-2-5p	22	0	0
+hsa-miR-25-5p	21	0	0
+hsa-miR-25-3p	22	0	0
+hsa-miR-26a-5p	22	0	0
+hsa-miR-26a-1-3p	22	0	0
+hsa-miR-26b-5p	21	0	0
+hsa-miR-26b-3p	22	0	0
+hsa-miR-27a-5p	22	0	0
+hsa-miR-27a-3p	21	0	0
+hsa-miR-28-5p	22	0	0
+hsa-miR-28-3p	22	0	0
+hsa-miR-29a-5p	22	0	0
+hsa-miR-29a-3p	22	0	0
+hsa-miR-30a-5p	22	0	0
+hsa-miR-30a-3p	22	0	0
+hsa-miR-31-5p	21	0	0
+hsa-miR-31-3p	22	0	0
+hsa-miR-32-5p	22	0	0
+hsa-miR-32-3p	22	0	0
+hsa-miR-33a-5p	21	0	0
+hsa-miR-33a-3p	22	0	0
+hsa-miR-92a-1-5p	23	0	0
+hsa-miR-92a-3p	22	0	0
+hsa-miR-92a-2-5p	22	0	0
+hsa-miR-93-5p	23	0	0
+hsa-miR-93-3p	22	0	0
+hsa-miR-95-5p	21	0	0
+hsa-miR-95-3p	22	0	0
+hsa-miR-96-5p	23	0	0
+hsa-miR-96-3p	22	0	0
+hsa-miR-98-5p	22	0	0
+hsa-miR-98-3p	22	0	0
+hsa-miR-99a-5p	22	0	0
+hsa-miR-99a-3p	22	0	0
+hsa-miR-100-5p	22	0	0
+hsa-miR-100-3p	22	0	0
+hsa-miR-101-5p	22	0	0
+hsa-miR-101-3p	21	0	0
+hsa-miR-29b-1-5p	24	0	0
+hsa-miR-29b-3p	23	0	0
+hsa-miR-29b-2-5p	22	0	0
+hsa-miR-103a-2-5p	23	0	0
+hsa-miR-103a-3p	23	0	0
+hsa-miR-105-5p	23	0	0
+hsa-miR-105-3p	22	0	0
+hsa-miR-106a-5p	23	0	0
+hsa-miR-106a-3p	22	0	0
+hsa-miR-107	23	0	0
+hsa-miR-16-2-3p	22	0	0
+hsa-miR-192-5p	21	0	0
+hsa-miR-192-3p	22	0	0
+hsa-miR-196a-5p	22	0	0
+hsa-miR-197-5p	23	0	0
+hsa-miR-197-3p	22	0	0
+hsa-miR-198	22	0	0
+hsa-miR-199a-5p	23	0	0
+hsa-miR-199a-3p	22	0	0
+hsa-miR-208a-5p	22	0	0
+hsa-miR-208a-3p	22	0	0
+hsa-miR-129-5p	21	0	0
+hsa-miR-129-1-3p	22	0	0
+hsa-miR-148a-5p	22	0	0
+hsa-miR-148a-3p	22	0	0
+hsa-miR-30c-5p	23	0	0
+hsa-miR-30c-2-3p	22	0	0
+hsa-miR-30d-5p	22	0	0
+hsa-miR-30d-3p	22	0	0
+hsa-miR-139-5p	23	0	0
+hsa-miR-139-3p	23	0	0
+hsa-miR-147a	20	0	0
+hsa-miR-7-5p	23	0	0
+hsa-miR-7-1-3p	22	0	0
+hsa-miR-7-2-3p	22	0	0
+hsa-miR-10a-5p	23	0	0
+hsa-miR-10a-3p	22	0	0
+hsa-miR-10b-5p	23	0	0
+hsa-miR-10b-3p	22	0	0
+hsa-miR-34a-5p	22	0	0
+hsa-miR-34a-3p	22	0	0
+hsa-miR-181a-5p	23	0	0
+hsa-miR-181a-2-3p	22	0	0
+hsa-miR-181b-5p	23	0	0
+hsa-miR-181b-3p	21	0	0
+hsa-miR-181c-5p	22	0	0
+hsa-miR-181c-3p	22	0	0
+hsa-miR-182-5p	24	0	0
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+hsa-miR-183-5p	22	0	0
+hsa-miR-183-3p	22	0	0
+hsa-miR-187-5p	22	0	0
+hsa-miR-187-3p	22	0	0
+hsa-miR-196a-3p	22	0	0
+hsa-miR-199b-5p	23	0	0
+hsa-miR-199b-3p	22	0	0
+hsa-miR-203a-5p	25	0	0
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+hsa-miR-214-3p	22	0	0
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+hsa-miR-218-2-3p	22	0	0
+hsa-miR-219a-5p	21	0	0
+hsa-miR-219a-1-3p	22	0	0
+hsa-miR-221-5p	22	0	0
+hsa-miR-221-3p	23	0	0
+hsa-miR-222-5p	22	0	0
+hsa-miR-222-3p	21	0	0
+hsa-miR-223-5p	22	0	0
+hsa-miR-223-3p	22	0	0
+hsa-miR-224-5p	21	0	0
+hsa-miR-224-3p	23	0	0
+hsa-miR-200b-5p	22	0	0
+hsa-miR-200b-3p	22	0	0
+hsa-let-7g-5p	22	0	0
+hsa-let-7g-3p	21	0	0
+hsa-let-7i-5p	22	0	0
+hsa-let-7i-3p	22	0	0
+hsa-miR-1-3p	22	0	0
+hsa-miR-15b-5p	22	0	0
+hsa-miR-15b-3p	22	0	0
+hsa-miR-23b-5p	22	0	0
+hsa-miR-23b-3p	21	0	0
+hsa-miR-27b-5p	22	0	0
+hsa-miR-27b-3p	21	0	0
+hsa-miR-30b-5p	22	0	0
+hsa-miR-30b-3p	22	0	0
+hsa-miR-122-5p	22	0	0
+hsa-miR-122-3p	22	0	0
+hsa-miR-124-5p	22	0	0
+hsa-miR-124-3p	20	0	0
+hsa-miR-125b-5p	22	0	0
+hsa-miR-125b-1-3p	22	0	0
+hsa-miR-128-1-5p	23	0	0
+hsa-miR-128-3p	21	0	0
+hsa-miR-130a-5p	22	0	0
+hsa-miR-130a-3p	22	0	0
+hsa-miR-132-5p	22	0	0
+hsa-miR-132-3p	22	0	0
+hsa-miR-133a-5p	22	0	0
+hsa-miR-133a-3p	22	0	0
+hsa-miR-135a-5p	23	0	0
+hsa-miR-135a-3p	22	0	0
+hsa-miR-137	23	0	0
+hsa-miR-138-5p	23	0	0
+hsa-miR-138-2-3p	22	0	0
+hsa-miR-140-5p	22	0	0
+hsa-miR-140-3p	21	0	0
+hsa-miR-141-5p	22	0	0
+hsa-miR-141-3p	22	0	0
+hsa-miR-142-5p	21	0	0
+hsa-miR-142-3p	23	0	0
+hsa-miR-143-5p	22	0	0
+hsa-miR-143-3p	21	0	0
+hsa-miR-144-5p	22	0	0
+hsa-miR-144-3p	20	0	0
+hsa-miR-145-5p	23	0	0
+hsa-miR-145-3p	22	0	0
+hsa-miR-152-5p	23	0	0
+hsa-miR-152-3p	21	0	0
+hsa-miR-153-3p	22	0	0
+hsa-miR-153-5p	22	0	0
+hsa-miR-191-5p	23	0	0
+hsa-miR-191-3p	22	0	0
+hsa-miR-9-5p	23	0	0
+hsa-miR-9-3p	22	0	0
+hsa-miR-125a-5p	24	0	0
+hsa-miR-125a-3p	22	0	0
+hsa-miR-125b-2-3p	22	0	0
+hsa-miR-126-5p	21	0	0
+hsa-miR-126-3p	22	0	0
+hsa-miR-127-5p	22	0	0
+hsa-miR-127-3p	22	0	0
+hsa-miR-129-2-3p	22	0	0
+hsa-miR-134-5p	22	0	0
+hsa-miR-134-3p	23	0	0
+hsa-miR-136-5p	23	0	0
+hsa-miR-136-3p	22	0	0
+hsa-miR-138-1-3p	22	0	0
+hsa-miR-146a-5p	22	0	0
+hsa-miR-146a-3p	22	0	0
+hsa-miR-149-5p	23	0	0
+hsa-miR-149-3p	21	0	0
+hsa-miR-150-5p	22	0	0
+hsa-miR-150-3p	22	0	0
+hsa-miR-154-5p	22	0	0
+hsa-miR-154-3p	22	0	0
+hsa-miR-184	22	0	0
+hsa-miR-185-5p	22	0	0
+hsa-miR-185-3p	22	0	0
+hsa-miR-186-5p	22	0	0
+hsa-miR-186-3p	22	0	0
+hsa-miR-188-5p	21	0	0
+hsa-miR-188-3p	21	0	0
+hsa-miR-190a-5p	22	0	0
+hsa-miR-190a-3p	21	0	0
+hsa-miR-193a-5p	22	0	0
+hsa-miR-193a-3p	22	0	0
+hsa-miR-194-5p	22	0	0
+hsa-miR-195-5p	2100	0	0
+hsa-miR-195-3p	2200	0	0
+hsa-miR-206	22	0	0
+hsa-miR-320a	22	0	0
+hsa-miR-200c-5p	22	0	0
+hsa-miR-200c-3p	23	0	0
+hsa-miR-1-5p	22	0	0
+hsa-miR-155-5p	23	0	0
+hsa-miR-155-3p	22	0	0
+hsa-miR-181b-2-3p	20	0	0
+hsa-miR-128-2-5p	23	0	0
+hsa-miR-194-3p	22	0	0
+hsa-miR-106b-5p	21	0	0
+hsa-miR-106b-3p	22	0	0
+hsa-miR-29c-5p	22	0	0
+hsa-miR-29c-3p	22	0	0
+hsa-miR-30c-1-3p	22	0	0
+hsa-miR-200a-5p	22	0	0
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+hsa-miR-302a-3p	23	0	0
+hsa-miR-219a-2-3p	22	0	0
+hsa-miR-34b-5p	23	0	0
+hsa-miR-34b-3p	22	0	0
+hsa-miR-34c-5p	23	0	0
+hsa-miR-34c-3p	22	0	0
+hsa-miR-299-5p	22	0	0
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+hsa-miR-301a-3p	23	0	0
+hsa-miR-99b-5p	22	0	0
+hsa-miR-99b-3p	22	0	0
+hsa-miR-296-5p	21	0	0
+hsa-miR-296-3p	22	0	0
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+hsa-miR-361-5p	22	0	0
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+hsa-miR-365a-5p	23	0	0
+hsa-miR-365a-3p	22	0	0
+hsa-miR-365b-5p	22	0	0
+hsa-miR-365b-3p	22	0	0
+hsa-miR-302b-5p	22	0	0
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+hsa-miR-383-3p	19	0	0
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+hsa-miR-328-5p	23	0	0
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+hsa-miR-342-5p	21	0	0
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+hsa-miR-337-5p	21	0	0
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+hsa-miR-6814-3p	22	0	0
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+hsa-miR-6815-3p	21	0	0
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+hsa-miR-6816-3p	18	0	0
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+hsa-miR-6840-5p	24	0	0
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+hsa-miR-6852-5p	21	0	0
+hsa-miR-6852-3p	17	0	0
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+hsa-miR-6854-5p	24	0	0
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+hsa-miR-6855-5p	25	0	0
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+hsa-miR-6857-5p	22	0	0
+hsa-miR-6857-3p	21	0	0
+hsa-miR-6858-5p	22	0	0
+hsa-miR-6858-3p	22	0	0
+hsa-miR-6859-5p	23	0	0
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+hsa-miR-6883-3p	22	0	0
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+hsa-miR-6884-3p	23	0	0
+hsa-miR-6885-5p	25	0	0
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+hsa-miR-6886-5p	21	0	0
+hsa-miR-6886-3p	21	0	0
+hsa-miR-6887-5p	23	0	0
+hsa-miR-6887-3p	21	0	0
+hsa-miR-6888-5p	20	0	0
+hsa-miR-6888-3p	21	0	0
+hsa-miR-6889-5p	23	0	0
+hsa-miR-6889-3p	20	0	0
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+hsa-miR-7107-3p	27	0	0
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+hsa-miR-7110-3p	22	0	0
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+hsa-miR-8080	23	0	0
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+hsa-miR-9500	19	0	0
+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	670172
diff --git a/testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..89bd5773
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
+hsa-let-7a-1	80	123	0
+hsa-let-7a-2	72	169	0
+hsa-let-7a-3	74	6	0
+hsa-let-7b	83	116	0
+hsa-let-7c	84	60	0
+hsa-let-7d	87	6	0
+hsa-let-7e	79	22	0
+hsa-let-7f-1	87	155	0
+hsa-let-7f-2	83	28	0
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+hsa-mir-16-1	89	5	0
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+hsa-mir-18a	71	0	0
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+hsa-mir-19b-1	87	1	0
+hsa-mir-19b-2	96	1	0
+hsa-mir-20a	71	16	0
+hsa-mir-21	72	10	0
+hsa-mir-22	85	1	0
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+hsa-mir-24-2	73	0	0
+hsa-mir-25	84	8	0
+hsa-mir-26a-1	77	35	0
+hsa-mir-26b	77	5	0
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+hsa-mir-28	86	4	0
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+hsa-mir-30a	71	42	0
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+hsa-mir-92a-1	78	29	0
+hsa-mir-92a-2	75	35	0
+hsa-mir-93	80	46	0
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+hsa-mir-96	78	0	0
+hsa-mir-98	119	10	0
+hsa-mir-99a	81	20	0
+hsa-mir-100	80	4	0
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diff --git a/testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats b/testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats
new file mode 100644
index 00000000..ab72cc01
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats
@@ -0,0 +1,2589 @@
+hsa-let-7a-5p	22	0	0
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+hsa-miR-20a-3p	22	0	0
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+hsa-miR-6875-5p	21	0	0
+hsa-miR-6875-3p	22	0	0
+hsa-miR-6876-5p	22	0	0
+hsa-miR-6876-3p	24	0	0
+hsa-miR-6877-5p	22	0	0
+hsa-miR-6877-3p	21	0	0
+hsa-miR-6878-5p	23	0	0
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+hsa-miR-6879-5p	22	0	0
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+hsa-miR-6880-3p	21	0	0
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+hsa-miR-6885-5p	25	0	0
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+hsa-miR-6887-5p	23	0	0
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+hsa-miR-6889-5p	23	0	0
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+hsa-miR-6890-5p	21	0	0
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+hsa-miR-9500	19	0	0
+hsa-miR-548bb-5p	22	0	0
+hsa-miR-548bb-3p	22	0	0
+*	0	0	2690903
diff --git a/testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats
new file mode 100644
index 00000000..9e2fa443
--- /dev/null
+++ b/testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats
@@ -0,0 +1,1882 @@
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+hsa-mir-8055	97	0	0
+hsa-mir-8056	82	0	0
+hsa-mir-8057	69	0	0
+hsa-mir-8058	89	0	0
+hsa-mir-8059	81	0	0
+hsa-mir-8060	76	0	0
+hsa-mir-8061	75	0	0
+hsa-mir-8062	85	0	0
+hsa-mir-8063	81	0	0
+hsa-mir-8064	90	0	0
+hsa-mir-8065	100	0	0
+hsa-mir-8066	78	0	0
+hsa-mir-8067	77	0	0
+hsa-mir-8068	68	0	0
+hsa-mir-8069-1	86	0	0
+hsa-mir-8070	88	0	0
+hsa-mir-8071-1	65	0	0
+hsa-mir-8072	80	3	0
+hsa-mir-8073	72	0	0
+hsa-mir-8074	81	0	0
+hsa-mir-8075	80	0	0
+hsa-mir-8076	83	0	0
+hsa-mir-8077	75	3	0
+hsa-mir-8078	84	1	0
+hsa-mir-8079	72	0	0
+hsa-mir-8080	89	0	0
+hsa-mir-8081	95	0	0
+hsa-mir-8082	81	0	0
+hsa-mir-8083	89	0	0
+hsa-mir-8084	89	0	0
+hsa-mir-8085	65	0	0
+hsa-mir-8086	93	4	0
+hsa-mir-8087	78	0	0
+hsa-mir-8088	86	0	0
+hsa-mir-8089	82	5	0
+hsa-mir-6862-2	70	0	0
+hsa-mir-8071-2	65	0	0
+hsa-mir-6770-2	60	0	0
+hsa-mir-6770-3	60	0	0
+hsa-mir-6859-2	68	0	0
+hsa-mir-6859-3	68	0	0
+hsa-mir-8485	91	0	0
+hsa-mir-9500	65	0	0
+hsa-mir-548bb	66	0	0
+hsa-mir-3179-4	84	0	0
+hsa-mir-1244-4	85	0	0
+hsa-mir-3648-2	180	5	0
+hsa-mir-3670-3	65	0	0
+hsa-mir-3670-4	65	0	0
+hsa-mir-3687-2	61	0	0
+hsa-mir-6724-2	92	1	0
+hsa-mir-6724-3	92	1	0
+hsa-mir-6724-4	92	1	0
+hsa-mir-8069-2	86	0	0
+hsa-mir-941-5	72	0	0
+hsa-mir-6859-4	68	0	0
+hsa-mir-5701-3	82	0	0
+*	0	0	2651242
diff --git a/testdata/smrnaseq/design.txt b/testdata/smrnaseq/design.txt
new file mode 100644
index 00000000..4983798f
--- /dev/null
+++ b/testdata/smrnaseq/design.txt
@@ -0,0 +1 @@
+~ batch + condition_treatment
diff --git a/testdata/smrnaseq/requested_genes.txt b/testdata/smrnaseq/requested_genes.txt
new file mode 100644
index 00000000..341190be
--- /dev/null
+++ b/testdata/smrnaseq/requested_genes.txt
@@ -0,0 +1,5 @@
+hsa-mir-100
+hsa-mir-125b-1
+hsa-mir-200a
+hsa-mir-505
+hsa-let-7c
\ No newline at end of file
diff --git a/testdata/smrnaseq/samplesheet.tsv b/testdata/smrnaseq/samplesheet.tsv
new file mode 100644
index 00000000..ddd07d13
--- /dev/null
+++ b/testdata/smrnaseq/samplesheet.tsv
@@ -0,0 +1,9 @@
+QBiC Code	Condition: treatment	batch	Secondary Name
+Clone1_NN1	clone1	1	1
+Clone1_NN3	clone1	3	2
+Clone9_NN1	clone9	1	3
+Clone9_NN2	clone9	2	4
+Clone9_NN3	clone9	3	5
+Control_N1	control	1	6
+Control_N2	control	2	7
+Control_N3	control	3	8
diff --git a/testdata/smrnaseq/software_versions.yml b/testdata/smrnaseq/software_versions.yml
new file mode 100644
index 00000000..2c314188
--- /dev/null
+++ b/testdata/smrnaseq/software_versions.yml
@@ -0,0 +1,64 @@
+BOWTIE_MAP_GENOME:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_HAIRPIN:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_MATURE:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_SEQCLUSTER:
+  bowtie: 1.3.1
+  samtools: '1.14'
+CUSTOM_DUMPSOFTWAREVERSIONS:
+  python: 3.11.0
+  yaml: '6.0'
+FASTP:
+  fastp: 0.23.2
+FASTQC_RAW:
+  fastqc: 0.11.9
+FASTQC_TRIM:
+  fastqc: 0.11.9
+FORMAT_HAIRPIN:
+  fastx_toolkit: 0.0.14
+FORMAT_MATURE:
+  fastx_toolkit: 0.0.14
+INDEX_GENOME:
+  bowtie: 1.3.1
+INDEX_HAIRPIN:
+  bowtie: 1.3.1
+INDEX_MATURE:
+  bowtie: 1.3.1
+MIRDEEP2_MAPPER:
+  mapper: 2.0.1
+MIRDEEP2_PIGZ":
+  pigz: 2.3.4
+MIRDEEP2_RUN":
+  mirdeep2: 2.0.1
+MIRTOP_QUANT":
+  mirtop: 0.4.25
+MIRTRACE_RUN":
+  mirtrace: 1.0.1
+PARSE_HAIRPIN":
+  seqkit: 2.3.0
+PARSE_MATURE":
+  seqkit: 2.3.0
+SAMPLESHEET_CHECK:
+  python: 3.8.3
+SAMTOOLS_FLAGSTAT:
+  samtools: '1.17'
+SAMTOOLS_IDXSTATS:
+  samtools: '1.17'
+SAMTOOLS_INDEX:
+  samtools: '1.17'
+SAMTOOLS_SORT:
+  samtools: '1.17'
+SAMTOOLS_STATS:
+  samtools: '1.17'
+SEQCLUSTER_SEQUENCES":
+  seqcluster: 1.2.9
+TABLE_MERGE:
+  r-base: 3.6.2
+Workflow:
+  Nextflow: 22.10.2
+  nf-core/smrnaseq: 2.2.1

From 9f3149f99c024f68eb051464187154120ce67cd4 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 12:59:18 +0200
Subject: [PATCH 093/124] editorconfig

---
 assets/references.bibtex              | 316 +++++++++++++-------------
 testdata/smrnaseq/requested_genes.txt |   2 +-
 2 files changed, 159 insertions(+), 159 deletions(-)

diff --git a/assets/references.bibtex b/assets/references.bibtex
index b16e7e87..92bccdc4 100644
--- a/assets/references.bibtex
+++ b/assets/references.bibtex
@@ -1,95 +1,95 @@
 @article{ewels2016multiqc,
-    title={MultiQC: summarize analysis results for multiple tools and samples in a single report},
-    author={Ewels, Philip and Magnusson, M{\aa}ns and Lundin, Sverker and K{\"a}ller, Max},
-    journal={Bioinformatics},
-    volume={32},
-    number={19},
-    pages={3047--3048},
-    year={2016},
-    publisher={Oxford University Press}
+title={MultiQC: summarize analysis results for multiple tools and samples in a single report},
+author={Ewels, Philip and Magnusson, M{\aa}ns and Lundin, Sverker and K{\"a}ller, Max},
+journal={Bioinformatics},
+volume={32},
+number={19},
+pages={3047--3048},
+year={2016},
+publisher={Oxford University Press}
 }
 @article{kim2015hisat,
-    title={HISAT: a fast spliced aligner with low memory requirements},
-    author={Kim, Daehwan and Langmead, Ben and Salzberg, Steven L},
-    journal={Nature methods},
-    volume={12},
-    number={4},
-    pages={357},
-    year={2015},
-    publisher={Nature Publishing Group}
+title={HISAT: a fast spliced aligner with low memory requirements},
+author={Kim, Daehwan and Langmead, Ben and Salzberg, Steven L},
+journal={Nature methods},
+volume={12},
+number={4},
+pages={357},
+year={2015},
+publisher={Nature Publishing Group}
 }
 @article{wang2012rseqc,
-    title={RSeQC: quality control of RNA-seq experiments},
-    author={Wang, Liguo and Wang, Shengqin and Li, Wei},
-    journal={Bioinformatics},
-    volume={28},
-    number={16},
-    pages={2184--2185},
-    year={2012},
-    publisher={Oxford University Press}
+title={RSeQC: quality control of RNA-seq experiments},
+author={Wang, Liguo and Wang, Shengqin and Li, Wei},
+journal={Bioinformatics},
+volume={28},
+number={16},
+pages={2184--2185},
+year={2012},
+publisher={Oxford University Press}
 }
 @article{liao2014featurecounts,
-    title={featureCounts: an efficient general purpose program for assigning sequence reads to genomic features},
-    author={Liao, Yang and Smyth, Gordon K and Shi, Wei},
-    journal={Bioinformatics},
-    volume={30},
-    number={7},
-    pages={923--930},
-    year={2014}
+title={featureCounts: an efficient general purpose program for assigning sequence reads to genomic features},
+author={Liao, Yang and Smyth, Gordon K and Shi, Wei},
+journal={Bioinformatics},
+volume={30},
+number={7},
+pages={923--930},
+year={2014}
 }
 @article{love2014differential,
-    title={Differential analysis of count data--the DESeq2 package},
-    author={Love, Michael and Anders, Simon and Huber, Wolfgang},
-    journal={Genome Biol},
-    volume={15},
-    pages={550},
-    year={2014}
+title={Differential analysis of count data--the DESeq2 package},
+author={Love, Michael and Anders, Simon and Huber, Wolfgang},
+journal={Genome Biol},
+volume={15},
+pages={550},
+year={2014}
 }
 @article{Li2009,
-    author = {Li, Heng and Durbin, Richard},
-    title = {Fast and Accurate Short Read Alignment with Burrows--Wheeler Transform},
-    journal = {Bioinformatics},
-    issue_date = {July 2009},
-    volume = {25},
-    number = {14},
-    month = jul,
-    year = {2009},
-    issn = {1367-4803},
-    pages = {1754--1760},
-    numpages = {7},
-    url = {http://dx.doi.org/10.1093/bioinformatics/btp324},
-    doi = {10.1093/bioinformatics/btp324},
-    acmid = {1576130},
-    publisher = {Oxford University Press},
-    address = {Oxford, UK},
+author = {Li, Heng and Durbin, Richard},
+title = {Fast and Accurate Short Read Alignment with Burrows--Wheeler Transform},
+journal = {Bioinformatics},
+issue_date = {July 2009},
+volume = {25},
+number = {14},
+month = jul,
+year = {2009},
+issn = {1367-4803},
+pages = {1754--1760},
+numpages = {7},
+url = {http://dx.doi.org/10.1093/bioinformatics/btp324},
+doi = {10.1093/bioinformatics/btp324},
+acmid = {1576130},
+publisher = {Oxford University Press},
+address = {Oxford, UK},
 }
 @article{Lex2014,
-    doi = {10.1109/tvcg.2014.2346248},
-    url = {https://doi.org/10.1109/tvcg.2014.2346248},
-    year    = {2014},
-    month = {dec},
-    publisher = {Institute of Electrical and Electronics Engineers ({IEEE})},
-    volume = {20},
-    number = {12},
-    pages = {1983--1992},
-    author = {Alexander Lex and Nils Gehlenborg and Hendrik Strobelt and Romain Vuillemot and Hanspeter Pfister},
-    title = {{UpSet}: Visualization of Intersecting Sets},
-    journal = {{IEEE} Transactions on Visualization and Computer Graphics}
+doi = {10.1109/tvcg.2014.2346248},
+url = {https://doi.org/10.1109/tvcg.2014.2346248},
+year    = {2014},
+month = {dec},
+publisher = {Institute of Electrical and Electronics Engineers ({IEEE})},
+volume = {20},
+number = {12},
+pages = {1983--1992},
+author = {Alexander Lex and Nils Gehlenborg and Hendrik Strobelt and Romain Vuillemot and Hanspeter Pfister},
+title = {{UpSet}: Visualization of Intersecting Sets},
+journal = {{IEEE} Transactions on Visualization and Computer Graphics}
 }
 @article{Cingolani2012,
-    title={Using Drosophila melanogaster as a model for genotoxic chemical mutational studies with a new program, SnpSift},
-    author={Cingolani, P. and Patel, V.M. and Coon, M. and Nguyen, T. and Land, S.J. and Ruden, D.M. and Lu, X.},
-    journal={Frontiers in Genetics},
-    volume={3},
-    year={2012},
-    publisher={Frontiers Media SA}
+title={Using Drosophila melanogaster as a model for genotoxic chemical mutational studies with a new program, SnpSift},
+author={Cingolani, P. and Patel, V.M. and Coon, M. and Nguyen, T. and Land, S.J. and Ruden, D.M. and Lu, X.},
+journal={Frontiers in Genetics},
+volume={3},
+year={2012},
+publisher={Frontiers Media SA}
 }
 @article{Garrison2012,
-    title={Haplotype-based variant detection from short-read sequencing},
-    author={Garrison, E. and Marth, G.},
-    journal={arXiv:1207.3907},
-    publisher={[q-bio.GN]},
-    year={2012}
+title={Haplotype-based variant detection from short-read sequencing},
+author={Garrison, E. and Marth, G.},
+journal={arXiv:1207.3907},
+publisher={[q-bio.GN]},
+year={2012}
 }
 @article{Dobin2013,
 author = {Dobin, A. and Davis, C.A. and Schlesinger, F. and Drenkow, J. and Zaleski, C. and Jha, S. and Batut, P. and Chaisson, M. and Gingeras, T.R.},
@@ -168,109 +168,109 @@ title="dupRadar: a Bioconductor package for the assessment of PCR artifacts in R
 journal="BMC Bioinformatics",
 }
 @article{krueger2012trim,
-    title={Trim Galore: a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files, with some extra functionality for MspI-digested RRBS-type (Reduced Representation Bisufite-Seq) libraries},
-    author={Krueger, F},
-    journal={URL http://www.bioinformatics.babraham.ac.uk/projects/trim\_galore/},
-    year={2012}
+title={Trim Galore: a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files, with some extra functionality for MspI-digested RRBS-type (Reduced Representation Bisufite-Seq) libraries},
+author={Krueger, F},
+journal={URL http://www.bioinformatics.babraham.ac.uk/projects/trim\_galore/},
+year={2012}
 }
 @article{sayols2016dupradar,
-    title={dupRadar: a Bioconductor package for the assessment of PCR artifacts in RNA-Seq data},
-    author={Sayols, Sergi and Scherzinger, Denise and Klein, Holger},
-    journal={BMC bioinformatics},
-    volume={17},
-    number={1},
-    pages={428},
-    year={2016},
-    publisher={BioMed Central}
+title={dupRadar: a Bioconductor package for the assessment of PCR artifacts in RNA-Seq data},
+author={Sayols, Sergi and Scherzinger, Denise and Klein, Holger},
+journal={BMC bioinformatics},
+volume={17},
+number={1},
+pages={428},
+year={2016},
+publisher={BioMed Central}
 }
 @misc{andrews2010fastqc,
-    title={FastQC: a quality control tool for high throughput sequence data},
-    author={Andrews, Simon and others},
-    year={2010}
+title={FastQC: a quality control tool for high throughput sequence data},
+author={Andrews, Simon and others},
+year={2010}
 }
 @Book{ggplot,
-    author = {Hadley Wickham},
-    title = {ggplot2: Elegant Graphics for Data Analysis},
-    publisher = {Springer-Verlag New York},
-    year = {2016},
-    isbn = {978-3-319-24277-4},
-    url = {http://ggplot2.org},
+author = {Hadley Wickham},
+title = {ggplot2: Elegant Graphics for Data Analysis},
+publisher = {Springer-Verlag New York},
+year = {2016},
+isbn = {978-3-319-24277-4},
+url = {http://ggplot2.org},
 }
 @article{anders2012detecting,
-    title={Detecting differential usage of exons from RNA-seq data},
-    author={Anders, Simon and Reyes, Alejandro and Huber, Wolfgang},
-    journal={Genome research},
-    volume={22},
-    number={10},
-    pages={2008--2017},
-    year={2012},
-    publisher={Cold Spring Harbor Lab}
+title={Detecting differential usage of exons from RNA-seq data},
+author={Anders, Simon and Reyes, Alejandro and Huber, Wolfgang},
+journal={Genome research},
+volume={22},
+number={10},
+pages={2008--2017},
+year={2012},
+publisher={Cold Spring Harbor Lab}
 }
 @article{ewels2020nf,
-    title={The nf-core framework for community-curated bioinformatics pipelines},
-    author={Ewels, Philip A and Peltzer, Alexander and Fillinger, Sven and Patel, Harshil and Alneberg, Johannes and Wilm, Andreas and Garcia, Maxime Ulysse and Di Tommaso, Paolo and Nahnsen, Sven},
-    journal={Nature biotechnology},
-    volume={38},
-    number={3},
-    pages={276--278},
-    year={2020},
-    publisher={Nature Publishing Group}
+title={The nf-core framework for community-curated bioinformatics pipelines},
+author={Ewels, Philip A and Peltzer, Alexander and Fillinger, Sven and Patel, Harshil and Alneberg, Johannes and Wilm, Andreas and Garcia, Maxime Ulysse and Di Tommaso, Paolo and Nahnsen, Sven},
+journal={Nature biotechnology},
+volume={38},
+number={3},
+pages={276--278},
+year={2020},
+publisher={Nature Publishing Group}
 }
 @article{kang2018mirtrace,
-  title={miRTrace reveals the organismal origins of microRNA sequencing data},
-  author={Kang, Wenjing and Eldfjell, Yrin and Fromm, Bastian and Estivill, Xavier and Biryukova, Inna and Friedl{\"a}nder, Marc R},
-  journal={Genome biology},
-  volume={19},
-  number={1},
-  pages={1--15},
-  year={2018},
-  publisher={BioMed Central}
+title={miRTrace reveals the organismal origins of microRNA sequencing data},
+author={Kang, Wenjing and Eldfjell, Yrin and Fromm, Bastian and Estivill, Xavier and Biryukova, Inna and Friedl{\"a}nder, Marc R},
+journal={Genome biology},
+volume={19},
+number={1},
+pages={1--15},
+year={2018},
+publisher={BioMed Central}
 }
 @article{langmead2009ultrafast,
-  title={Ultrafast and memory-efficient alignment of short DNA sequences to the human genome},
-  author={Langmead, Ben and Trapnell, Cole and Pop, Mihai and Salzberg, Steven L},
-  journal={Genome biology},
-  volume={10},
-  number={3},
-  pages={1--10},
-  year={2009},
-  publisher={BioMed Central}
+title={Ultrafast and memory-efficient alignment of short DNA sequences to the human genome},
+author={Langmead, Ben and Trapnell, Cole and Pop, Mihai and Salzberg, Steven L},
+journal={Genome biology},
+volume={10},
+number={3},
+pages={1--10},
+year={2009},
+publisher={BioMed Central}
 }
 @article{chen2018fastp,
-  title={fastp: an ultra-fast all-in-one FASTQ preprocessor},
-  author={Chen, Shifu and Zhou, Yanqing and Chen, Yaru and Gu, Jia},
-  journal={Bioinformatics},
-  volume={34},
-  number={17},
-  pages={i884--i890},
-  year={2018},
-  publisher={Oxford University Press}
+title={fastp: an ultra-fast all-in-one FASTQ preprocessor},
+author={Chen, Shifu and Zhou, Yanqing and Chen, Yaru and Gu, Jia},
+journal={Bioinformatics},
+volume={34},
+number={17},
+pages={i884--i890},
+year={2018},
+publisher={Oxford University Press}
 }
 @article{chen2023ultrafast,
-  title={Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp},
-  author={Chen, Shifu},
-  journal={iMeta},
-  pages={e107},
-  year={2023},
-  publisher={Wiley Online Library}
+title={Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp},
+author={Chen, Shifu},
+journal={iMeta},
+pages={e107},
+year={2023},
+publisher={Wiley Online Library}
 }
 @article{li2009sequence,
-  title={The sequence alignment/map format and SAMtools},
-  author={Li, Heng and Handsaker, Bob and Wysoker, Alec and Fennell, Tim and Ruan, Jue and Homer, Nils and Marth, Gabor and Abecasis, Goncalo and Durbin, Richard and 1000 Genome Project Data Processing Subgroup},
-  journal={bioinformatics},
-  volume={25},
-  number={16},
-  pages={2078--2079},
-  year={2009},
-  publisher={Oxford University Press}
+title={The sequence alignment/map format and SAMtools},
+author={Li, Heng and Handsaker, Bob and Wysoker, Alec and Fennell, Tim and Ruan, Jue and Homer, Nils and Marth, Gabor and Abecasis, Goncalo and Durbin, Richard and 1000 Genome Project Data Processing Subgroup},
+journal={bioinformatics},
+volume={25},
+number={16},
+pages={2078--2079},
+year={2009},
+publisher={Oxford University Press}
 }
 @article{li2011statistical,
-  title={A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data},
-  author={Li, Heng},
-  journal={Bioinformatics},
-  volume={27},
-  number={21},
-  pages={2987--2993},
-  year={2011},
-  publisher={Oxford University Press}
+title={A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data},
+author={Li, Heng},
+journal={Bioinformatics},
+volume={27},
+number={21},
+pages={2987--2993},
+year={2011},
+publisher={Oxford University Press}
 }
diff --git a/testdata/smrnaseq/requested_genes.txt b/testdata/smrnaseq/requested_genes.txt
index 341190be..22e59997 100644
--- a/testdata/smrnaseq/requested_genes.txt
+++ b/testdata/smrnaseq/requested_genes.txt
@@ -2,4 +2,4 @@ hsa-mir-100
 hsa-mir-125b-1
 hsa-mir-200a
 hsa-mir-505
-hsa-let-7c
\ No newline at end of file
+hsa-let-7c

From bb2eb84a8b9ee2c0e629c0682a9caf23ca9ae13d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 14:04:46 +0200
Subject: [PATCH 094/124] undid an accidental change

---
 assets/RNAseq_report.Rmd | 10 ++++------
 1 file changed, 4 insertions(+), 6 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 4612c1e2..8d3d3135 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -524,9 +524,7 @@ DT::datatable(df_QA_neat, options = list(scrollX  = "100%", scrollY = "400px", p
 ################## RUN DESEQ2 ######################################
 # Run DESeq function
 if (params$input_type %in% c("featurecounts", "smrnaseq")) {
-
     cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]]))))
-
 } else if (params$input_type %in% c("rsem", "salmon")) {
     ## Create a dataframe which consists of both the gene id and the transcript name
     gtf <- rtracklayer::import(params$path_gtf)
@@ -1258,7 +1256,7 @@ if (isProvided(params$path_contrast_matrix)){
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1318,7 +1316,7 @@ if (isProvided(params$path_contrast_list)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <- DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1360,7 +1358,7 @@ if (isProvided(params$path_contrast_pairs)) {
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <-  DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <-  subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
 
         # Round table before writing
@@ -1401,7 +1399,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <-  DE_genes_contrast_genename #subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <-  subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {

From 3452abd158ed3003d6ea84304531f6f3ad3b6c37 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 14:58:12 +0200
Subject: [PATCH 095/124] bugfix

---
 tests/test_smrnaseq.yml | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml
index 493b7b36..31beedd6 100644
--- a/tests/test_smrnaseq.yml
+++ b/tests/test_smrnaseq.yml
@@ -1,7 +1,7 @@
 - name: Run test profile with smrnaseq
   command: nextflow run main.nf -profile test_smrnaseq,docker --citest true --round_DE 3
   tags:
-    - test_star_salmon
+    - test_smrnaseq
   files:
     - path: results_test/differential_gene_expression/metadata/smrnaseq_files.txt
       md5sum: ce3cc117fc953a8ff4d17d0118123faf

From 2767b05312931142c1eb0a8bb8532facdf225b21 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 14:59:50 +0200
Subject: [PATCH 096/124] fixed test data path

---
 conf/test_smrnaseq.config | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config
index e421a1c3..f69cfaab 100644
--- a/conf/test_smrnaseq.config
+++ b/conf/test_smrnaseq.config
@@ -17,7 +17,7 @@ params {
     // Input data
     input = 'testdata/smrnaseq/samplesheet.tsv'
     gene_counts = 'testdata/smrnaseq/counts'
-    model = '/home-link/iivow01/git/rnadeseq/testdata/smrnaseq/design.txt'
+    model = 'testdata/smrnaseq/design.txt'
     logFC_threshold = 1
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
     software_versions = '/home-link/iivow01/git/rnadeseq/testdata/smrnaseq/software_versions.yml'

From 82ff93b83ac02a8f9fc0b72a5f984eb736e15c3b Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 15:12:14 +0200
Subject: [PATCH 097/124] fixed another path

---
 conf/test_smrnaseq.config | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config
index f69cfaab..ac69f9eb 100644
--- a/conf/test_smrnaseq.config
+++ b/conf/test_smrnaseq.config
@@ -20,7 +20,7 @@ params {
     model = 'testdata/smrnaseq/design.txt'
     logFC_threshold = 1
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
-    software_versions = '/home-link/iivow01/git/rnadeseq/testdata/smrnaseq/software_versions.yml'
+    software_versions = 'testdata/smrnaseq/software_versions.yml'
     genelist = 'testdata/smrnaseq/requested_genes.txt'
     input_type = 'smrnaseq'
     run_pathway_analysis = true

From abad0ff3c257375b403f426224a7b89e0ec48efd Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 15:30:28 +0200
Subject: [PATCH 098/124] changed path of software versions

---
 conf/test_smrnaseq.config               |  2 +-
 testdata/software_versions_smrnaseq.yml | 64 +++++++++++++++++++++++++
 2 files changed, 65 insertions(+), 1 deletion(-)
 create mode 100644 testdata/software_versions_smrnaseq.yml

diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config
index ac69f9eb..797fa332 100644
--- a/conf/test_smrnaseq.config
+++ b/conf/test_smrnaseq.config
@@ -20,7 +20,7 @@ params {
     model = 'testdata/smrnaseq/design.txt'
     logFC_threshold = 1
     project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv'
-    software_versions = 'testdata/smrnaseq/software_versions.yml'
+    software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_smrnaseq.yml'
     genelist = 'testdata/smrnaseq/requested_genes.txt'
     input_type = 'smrnaseq'
     run_pathway_analysis = true
diff --git a/testdata/software_versions_smrnaseq.yml b/testdata/software_versions_smrnaseq.yml
new file mode 100644
index 00000000..2c314188
--- /dev/null
+++ b/testdata/software_versions_smrnaseq.yml
@@ -0,0 +1,64 @@
+BOWTIE_MAP_GENOME:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_HAIRPIN:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_MATURE:
+  bowtie: 1.3.1
+  samtools: '1.14'
+BOWTIE_MAP_SEQCLUSTER:
+  bowtie: 1.3.1
+  samtools: '1.14'
+CUSTOM_DUMPSOFTWAREVERSIONS:
+  python: 3.11.0
+  yaml: '6.0'
+FASTP:
+  fastp: 0.23.2
+FASTQC_RAW:
+  fastqc: 0.11.9
+FASTQC_TRIM:
+  fastqc: 0.11.9
+FORMAT_HAIRPIN:
+  fastx_toolkit: 0.0.14
+FORMAT_MATURE:
+  fastx_toolkit: 0.0.14
+INDEX_GENOME:
+  bowtie: 1.3.1
+INDEX_HAIRPIN:
+  bowtie: 1.3.1
+INDEX_MATURE:
+  bowtie: 1.3.1
+MIRDEEP2_MAPPER:
+  mapper: 2.0.1
+MIRDEEP2_PIGZ":
+  pigz: 2.3.4
+MIRDEEP2_RUN":
+  mirdeep2: 2.0.1
+MIRTOP_QUANT":
+  mirtop: 0.4.25
+MIRTRACE_RUN":
+  mirtrace: 1.0.1
+PARSE_HAIRPIN":
+  seqkit: 2.3.0
+PARSE_MATURE":
+  seqkit: 2.3.0
+SAMPLESHEET_CHECK:
+  python: 3.8.3
+SAMTOOLS_FLAGSTAT:
+  samtools: '1.17'
+SAMTOOLS_IDXSTATS:
+  samtools: '1.17'
+SAMTOOLS_INDEX:
+  samtools: '1.17'
+SAMTOOLS_SORT:
+  samtools: '1.17'
+SAMTOOLS_STATS:
+  samtools: '1.17'
+SEQCLUSTER_SEQUENCES":
+  seqcluster: 1.2.9
+TABLE_MERGE:
+  r-base: 3.6.2
+Workflow:
+  Nextflow: 22.10.2
+  nf-core/smrnaseq: 2.2.1

From fccbb4c2374adea49ac45776852d1a8c5290b610 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 15:31:40 +0200
Subject: [PATCH 099/124] deleted old smrnaseq versions

---
 testdata/smrnaseq/software_versions.yml | 64 -------------------------
 1 file changed, 64 deletions(-)
 delete mode 100644 testdata/smrnaseq/software_versions.yml

diff --git a/testdata/smrnaseq/software_versions.yml b/testdata/smrnaseq/software_versions.yml
deleted file mode 100644
index 2c314188..00000000
--- a/testdata/smrnaseq/software_versions.yml
+++ /dev/null
@@ -1,64 +0,0 @@
-BOWTIE_MAP_GENOME:
-  bowtie: 1.3.1
-  samtools: '1.14'
-BOWTIE_MAP_HAIRPIN:
-  bowtie: 1.3.1
-  samtools: '1.14'
-BOWTIE_MAP_MATURE:
-  bowtie: 1.3.1
-  samtools: '1.14'
-BOWTIE_MAP_SEQCLUSTER:
-  bowtie: 1.3.1
-  samtools: '1.14'
-CUSTOM_DUMPSOFTWAREVERSIONS:
-  python: 3.11.0
-  yaml: '6.0'
-FASTP:
-  fastp: 0.23.2
-FASTQC_RAW:
-  fastqc: 0.11.9
-FASTQC_TRIM:
-  fastqc: 0.11.9
-FORMAT_HAIRPIN:
-  fastx_toolkit: 0.0.14
-FORMAT_MATURE:
-  fastx_toolkit: 0.0.14
-INDEX_GENOME:
-  bowtie: 1.3.1
-INDEX_HAIRPIN:
-  bowtie: 1.3.1
-INDEX_MATURE:
-  bowtie: 1.3.1
-MIRDEEP2_MAPPER:
-  mapper: 2.0.1
-MIRDEEP2_PIGZ":
-  pigz: 2.3.4
-MIRDEEP2_RUN":
-  mirdeep2: 2.0.1
-MIRTOP_QUANT":
-  mirtop: 0.4.25
-MIRTRACE_RUN":
-  mirtrace: 1.0.1
-PARSE_HAIRPIN":
-  seqkit: 2.3.0
-PARSE_MATURE":
-  seqkit: 2.3.0
-SAMPLESHEET_CHECK:
-  python: 3.8.3
-SAMTOOLS_FLAGSTAT:
-  samtools: '1.17'
-SAMTOOLS_IDXSTATS:
-  samtools: '1.17'
-SAMTOOLS_INDEX:
-  samtools: '1.17'
-SAMTOOLS_SORT:
-  samtools: '1.17'
-SAMTOOLS_STATS:
-  samtools: '1.17'
-SEQCLUSTER_SEQUENCES":
-  seqcluster: 1.2.9
-TABLE_MERGE:
-  r-base: 3.6.2
-Workflow:
-  Nextflow: 22.10.2
-  nf-core/smrnaseq: 2.2.1

From ef2ab181b4e921c1469aa367b741054acc4ce653 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 15:43:12 +0200
Subject: [PATCH 100/124] removed newline to open new pr

---
 assets/RNAseq_report.Rmd | 1 -
 1 file changed, 1 deletion(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 8d3d3135..73981782 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1644,7 +1644,6 @@ cat("Plots for the [genes of interest](./differential_gene_expression/metadata/g
 
 <!-- In case KEGG analysis was also included; differentiating the cases with and without contrasts -->
 
-
 <!-- PA start -->
 ```{r load_PA_libs, eval=params$pathway_analysis, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'}
 

From 9e4b43889c452b143b7224c8a3c9f18ccfb431e2 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 7 Aug 2023 15:57:39 +0200
Subject: [PATCH 101/124] updated changelog and md5sums

---
 CHANGELOG.md            |  3 ++-
 tests/test_smrnaseq.yml | 12 ++++++------
 2 files changed, 8 insertions(+), 7 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index e07a79a6..2c01d96f 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#213](https://github.com/qbic-pipelines/rnadeseq/pull/21) Added smrnaseq input support
+- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works
+- [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
 - [#202](https://github.com/qbic-pipelines/rnadeseq/pull/202) Added background list to pathway analysis
diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml
index 31beedd6..506bf6cf 100644
--- a/tests/test_smrnaseq.yml
+++ b/tests/test_smrnaseq.yml
@@ -6,9 +6,9 @@
     - path: results_test/differential_gene_expression/metadata/smrnaseq_files.txt
       md5sum: ce3cc117fc953a8ff4d17d0118123faf
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_clone9_vs_clone1.tsv
-      md5sum: 46ca6c2ec1046e088853c96b28cd30e3
+      md5sum: 9526e0b2be54b49a8d1694b6cdd939b7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_control_vs_clone1.tsv
-      md5sum: 74214d33037aeded1bd0c839973a9e4c
+      md5sum: e5e4bfab55e82a0d3d80b219d60f2df0
     - path: results_test/differential_gene_expression/final_gene_table/final_DE_gene_list.tsv
       md5sum: 9f4950203e56a94235b00a1d554fa78c
     - path: results_test/differential_gene_expression/gene_counts_tables/deseq2_library_scaled_gene_counts.tsv
@@ -76,14 +76,14 @@
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_results.tsv
-      md5sum: b5c4c68a304f08a54b2b9d8969334e8c
+      md5sum: 2637f5f6f4e74e7723280449e3ac647b
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_pathway_enrichment_results.tsv
-      md5sum: ab088ff01a4e3991be8a424e676afcdf
+      md5sum: a091e2ae9a678599d9e994082907d646
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.pdf
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.png
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.svg
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_results.tsv
-      md5sum: 4a2ed390ae3e9fea628bd9c014e88cc6
+      md5sum: 497cbe026880c9ea78c5591c7c4d07f7
     - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_pathway_enrichment_results.tsv
-      md5sum: db2149ba5f2dbb6d4aed3f6284072f39
+      md5sum: 63a2adf7c4a971dc7443503cf968e455
     - path: results_test/RNAseq_report.html

From b0bcc5bddf3156c28719dd95352d5c5279c3a35f Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 8 Aug 2023 14:04:19 +0200
Subject: [PATCH 102/124] cleanup from previous PR

---
 assets/RNAseq_report.Rmd | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 73981782..3dfde2d2 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -2255,8 +2255,8 @@ cat("No software versions file was provided for primary analysis.")
 ```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)}
 
 # In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is clearer in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
-# TODO remove F!
-if (params$input_type == "featurecounts" & F){
+
+if (params$input_type == "featurecounts"){
     cat(paste0(
     "## RNAseq data analysis
 

From a445369525be3cae48308dbe3a9fbd6c537c4100 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 14 Aug 2023 13:14:33 +0200
Subject: [PATCH 103/124] Updated container, fixed resulting bug in
 include_graphics, changed smrnaseq testdata with QBiC Code

---
 CHANGELOG.md                                  |  2 +-
 assets/RNAseq_report.Rmd                      | 26 ++++----
 environment.yml                               | 62 +++++++++----------
 ...tats => QABCD00001_mature.sorted.idxstats} |  0
 ...QABCD00001_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00002_mature.sorted.idxstats} |  0
 ...QABCD00002_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00003_mature.sorted.idxstats} |  0
 ...QABCD00003_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00004_mature.sorted.idxstats} |  0
 ...QABCD00004_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00005_mature.sorted.idxstats} |  0
 ...QABCD00005_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00006_mature.sorted.idxstats} |  0
 ...QABCD00006_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00007_mature.sorted.idxstats} |  0
 ...QABCD00007_mature_hairpin.sorted.idxstats} |  0
 ...tats => QABCD00008_mature.sorted.idxstats} |  0
 ...QABCD00008_mature_hairpin.sorted.idxstats} |  0
 testdata/smrnaseq/samplesheet.tsv             | 16 ++---
 20 files changed, 55 insertions(+), 51 deletions(-)
 rename testdata/smrnaseq/counts/{Clone1_NN1_mature.sorted.idxstats => QABCD00001_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone1_NN1_mature_hairpin.sorted.idxstats => QABCD00001_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone1_NN3_mature.sorted.idxstats => QABCD00002_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone1_NN3_mature_hairpin.sorted.idxstats => QABCD00002_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN1_mature.sorted.idxstats => QABCD00003_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN1_mature_hairpin.sorted.idxstats => QABCD00003_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN2_mature.sorted.idxstats => QABCD00004_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN2_mature_hairpin.sorted.idxstats => QABCD00004_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN3_mature.sorted.idxstats => QABCD00005_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Clone9_NN3_mature_hairpin.sorted.idxstats => QABCD00005_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N1_mature.sorted.idxstats => QABCD00006_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N1_mature_hairpin.sorted.idxstats => QABCD00006_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N2_mature.sorted.idxstats => QABCD00007_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N2_mature_hairpin.sorted.idxstats => QABCD00007_mature_hairpin.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N3_mature.sorted.idxstats => QABCD00008_mature.sorted.idxstats} (100%)
 rename testdata/smrnaseq/counts/{Control_N3_mature_hairpin.sorted.idxstats => QABCD00008_mature_hairpin.sorted.idxstats} (100%)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 2c01d96f..60fb4344 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works
+- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 3dfde2d2..59ea7c81 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -104,13 +104,14 @@ build_smrnaseq_df <- function(files) {
             if (rownames(file_tab) != rownames(merged_mat)) {
                 stop(paste("Rownames of smrnaseq file", file_tab, "differ from the rownames of the other files; exiting..."))
             }
-            merged_mat[[basename(f)]] <- file_tab[,2]
+            # Create column from QBiC code of current file name (substring of length 10)
+            merged_mat[[substr(basename(f), 1, 10)]] <- file_tab[,2]
         } else {
 
-            # Otherwise initiate it from third matrix column and rownames
+            # Otherwise initiate it from third matrix column and rownames; as above, only use QBiC code, not full file name
             merged_mat <- data.frame(file_tab[,2])
             rownames(merged_mat) <- rownames(file_tab)
-            colnames(merged_mat) <- c(basename(f))
+            colnames(merged_mat) <- c(substr(basename(f), 1, 10))
         }
     }
     merged_mat
@@ -941,7 +942,9 @@ The read mapping statistics to the reference genome for each sample are shown be
 ```
 
 ```{r STAR_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1800, fig.cap="STAR: Mapping Statistics", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"))
+# Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every call to not fail with some stupid error
+knitr.graphics.rel_path = F
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"), error=F)
 ```
 
 ```{r STAR_readnums,echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -953,7 +956,7 @@ cat(paste0("***
 ```
 
 ```{r STAR_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1800, fig.cap="STAR: Mapping Statistics", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1.svg"))
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1.svg"), error=F)
 ```
 
 ```{r FC_read_assignment, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -979,7 +982,7 @@ The statistics of read assignment to genes are shown below. Most reads should be
 ```
 
 ```{r FC_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="featureCounts: Assignments", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1_pc.svg"))
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1_pc.svg"), error=F)
 ```
 
 ```{r FC_readnums, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -993,7 +996,7 @@ cat(paste0("***
 ```
 
 ```{r FC_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="featureCounts: Assignments", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1.svg"))
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1.svg"), error=F)
 ```
 
 ```{r read_feat_distrib, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -1013,7 +1016,7 @@ In RNAseq experiments, the majority of the reads should be assigned to CDS exons
 ```
 
 ```{r RSeqC_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="RSeQC: Read Distribution", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_read_distribution_plot_1_pc.svg"))
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_read_distribution_plot_1_pc.svg"), error=F)
 ```
 
 ```{r RSeqC_readnums, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -1027,7 +1030,7 @@ cat(paste0("***
 ```
 
 ```{r RSeqC_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="RSeQC: Read Distribution", fig.align='center'}
-knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_read_distribution_plot_1.svg"))
+knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_read_distribution_plot_1.svg"), error=F)
 ```
 
 ```{r MQC_end, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))}
@@ -1667,7 +1670,7 @@ short_organism_name <- substr(organism,1,3)
 if (!require("BiocManager", quietly = TRUE)) {
     install.packages("BiocManager")
 }
-BiocManager::install(params$species_library, lib=species_dir, version="3.14", force=T)
+BiocManager::install(params$species_library, lib=species_dir, version="3.17", force=T)
 library(params$species_library, lib.loc=species_dir, character.only=T)
 species_library_installed <- get(params$species_library)
 
@@ -2303,7 +2306,8 @@ quant_tool,
 quant_version,
 "` ",
 quant_cite,
-"."
+".",
+"\nThe output from `SAMtools`, more specifically the  `<QBiC-Code>_mature_hairpin.sorted.idxstats` and `<QBiC-Code>_mature.sorted.idxstats` files, were then used to create a counts matrix. This was done by combining all of these files into a single table, using only the rownames and the third column (second column after the rownames) of each file which contains the counts. The last row of each file (which always has an asterisk `*` as rowname) was excluded. The resulting counts matrix was used as input for the `rnadeseq` pipeline."
 ))
 }
 ```
diff --git a/environment.yml b/environment.yml
index 3c62aa8a..57e1e3f1 100644
--- a/environment.yml
+++ b/environment.yml
@@ -8,40 +8,40 @@ channels:
   - defaults
   - r
 dependencies:
-  - bioconda::bioconductor-annotationdbi=1.56.2
-  - bioconda::bioconductor-deseq2=1.34.0
-  - bioconda::bioconductor-genefilter=1.76.0
-  - bioconda::bioconductor-genomeinfodbdata=1.2.7
-  - bioconda::bioconductor-impute=1.68.0
-  - bioconda::bioconductor-limma=3.50.3
-  - bioconda::bioconductor-pathview=1.34.0
-  - bioconda::bioconductor-rtracklayer=1.54.0
-  - bioconda::bioconductor-summarizedexperiment=1.24.0
-  - bioconda::bioconductor-tximeta=1.12.0
-  - bioconda::bioconductor-tximport=1.22.0
-  - bioconda::bioconductor-vsn=3.62.0
-  - conda-forge::pandoc=2.17.1.1
-  - conda-forge::r-base=4.1.2
+  - bioconda::bioconductor-annotationdbi=1.62.2
+  - bioconda::bioconductor-deseq2=1.40.2
+  - bioconda::bioconductor-genefilter=1.82.1
+  - bioconda::bioconductor-genomeinfodbdata=1.2.10
+  - bioconda::bioconductor-impute=1.74.1
+  - bioconda::bioconductor-limma=3.56.2
+  - bioconda::bioconductor-pathview=1.40.0
+  - bioconda::bioconductor-rtracklayer=1.60.0
+  - bioconda::bioconductor-summarizedexperiment=1.30.2
+  - bioconda::bioconductor-tximeta=1.18.0
+  - bioconda::bioconductor-tximport=1.28.0
+  - bioconda::bioconductor-vsn=3.68.0
+  - conda-forge::pandoc=3.1.3
+  - conda-forge::r-base=4.3.1
   - conda-forge::r-details=0.3.0
-  - conda-forge::r-dplyr=1.0.10
-  - conda-forge::r-dt=0.20
-  - conda-forge::r-extrafont=0.17
+  - conda-forge::r-dplyr=1.1.2
+  - conda-forge::r-dt=0.28
+  - conda-forge::r-extrafont=0.19
   - conda-forge::r-formattable=0.2.1
-  - conda-forge::r-ggplot2=3.3.5
-  - conda-forge::r-ggrepel=0.9.2
-  - conda-forge::r-ggvenn=0.1.9
-  - conda-forge::r-gplots=3.1.1
-  - conda-forge::r-gprofiler2=0.2.1
-  - conda-forge::r-heatmaply=1.3.0
+  - conda-forge::r-ggplot2=3.4.2
+  - conda-forge::r-ggrepel=0.9.3
+  - conda-forge::r-ggvenn=0.1.10
+  - conda-forge::r-gplots=3.1.3
+  - conda-forge::r-gprofiler2=0.2.2
+  - conda-forge::r-heatmaply=1.4.2
   - conda-forge::r-kableextra=1.3.4
-  - conda-forge::r-knitr=1.37
-  - conda-forge::r-optparse=1.7.1
+  - conda-forge::r-knitr=1.43
+  - conda-forge::r-optparse=1.7.3
   - conda-forge::r-pheatmap=1.0.12
-  - conda-forge::r-plyr=1.8.6
-  - conda-forge::r-rcolorbrewer=1.1_2
+  - conda-forge::r-plyr=1.8.8
+  - conda-forge::r-rcolorbrewer=1.1_3
   - conda-forge::r-reshape2=1.4.4
-  - conda-forge::r-rmarkdown=2.11
+  - conda-forge::r-rmarkdown=2.23
   - conda-forge::r-sessioninfo=1.2.2
-  - conda-forge::r-svglite=2.1.0
-  - conda-forge::r-tidyverse=1.3.1
-  - conda-forge::r-yaml=2.2.2
+  - conda-forge::r-svglite=2.1.1
+  - conda-forge::r-tidyverse=2.0.0
+  - conda-forge::r-yaml=2.3.7
\ No newline at end of file
diff --git a/testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00001_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone1_NN1_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00001_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00001_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone1_NN1_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00001_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00002_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone1_NN3_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00002_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00002_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone1_NN3_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00002_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00003_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN1_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00003_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00003_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN1_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00003_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00004_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN2_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00004_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00004_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN2_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00004_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00005_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN3_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00005_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00005_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Clone9_NN3_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00005_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00006_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N1_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00006_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00006_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N1_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00006_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00007_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N2_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00007_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00007_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N2_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00007_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00008_mature.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N3_mature.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00008_mature.sorted.idxstats
diff --git a/testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats b/testdata/smrnaseq/counts/QABCD00008_mature_hairpin.sorted.idxstats
similarity index 100%
rename from testdata/smrnaseq/counts/Control_N3_mature_hairpin.sorted.idxstats
rename to testdata/smrnaseq/counts/QABCD00008_mature_hairpin.sorted.idxstats
diff --git a/testdata/smrnaseq/samplesheet.tsv b/testdata/smrnaseq/samplesheet.tsv
index ddd07d13..12dc0507 100644
--- a/testdata/smrnaseq/samplesheet.tsv
+++ b/testdata/smrnaseq/samplesheet.tsv
@@ -1,9 +1,9 @@
 QBiC Code	Condition: treatment	batch	Secondary Name
-Clone1_NN1	clone1	1	1
-Clone1_NN3	clone1	3	2
-Clone9_NN1	clone9	1	3
-Clone9_NN2	clone9	2	4
-Clone9_NN3	clone9	3	5
-Control_N1	control	1	6
-Control_N2	control	2	7
-Control_N3	control	3	8
+QABCD00001	clone1	1	1
+QABCD00002	clone1	3	2
+QABCD00003	clone9	1	3
+QABCD00004	clone9	2	4
+QABCD00005	clone9	3	5
+QABCD00006	control	1	6
+QABCD00007	control	2	7
+QABCD00008	control	3	8

From 080e36b4f20c3ac8480d3ffddaedad61184f85a6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 14 Aug 2023 15:18:09 +0200
Subject: [PATCH 104/124] bugfix; made heatmap order equal between files in zip
 and plot in report

---
 CHANGELOG.md             | 2 +-
 assets/RNAseq_report.Rmd | 7 ++++---
 2 files changed, 5 insertions(+), 4 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 60fb4344..9d2edb00 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics
+- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics. Heatmaps are now equally ordered in zip and report
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 59ea7c81..05a416a3 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -101,7 +101,7 @@ build_smrnaseq_df <- function(files) {
         if (is.data.frame(merged_mat)) {
 
             # Check if rownames are the same
-            if (rownames(file_tab) != rownames(merged_mat)) {
+            if (!identical(rownames(file_tab), rownames(merged_mat))) {
                 stop(paste("Rownames of smrnaseq file", file_tab, "differ from the rownames of the other files; exiting..."))
             }
             # Create column from QBiC code of current file name (substring of length 10)
@@ -1146,10 +1146,11 @@ The original plot can be downloaded [here](./differential_gene_expression/plots/
 ##################  SAMPLE DISTANCES HEATMAP ##################
 # Sample distances, this chunk is hidden as otherwise, the heatmap is shown multiple times
 sampleDists <- dist(t(assay(if (run_rlog) rld else vsd)))
+sampleDistClust <- hclust(sampleDists)  # Do clustering as input for heatmaps so that their column orders are the same
 sampleDistMatrix <- as.matrix(sampleDists)
 colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255)
 par(oma=c(3,3,3,3))
-dist_heatmap <- pheatmap(mat = sampleDistMatrix, color=colors)
+dist_heatmap <- pheatmap(mat = sampleDistMatrix, color=colors, cluster_rows = sampleDistClust, cluster_cols = sampleDistClust)
 
 svg("differential_gene_expression/plots/Heatmaps_of_distances.svg")
 dist_heatmap
@@ -1162,7 +1163,7 @@ dist_heatmap
 dev.off()
 
 # Recompute heatmap with heatmaply for interactive plot in report
-dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey")
+dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey", Rowv = sampleDistClust, Colv = "Rowv")
 ```
 
 ```{r heatmap_show, echo=FALSE, message=FALSE, warning=FALSE, out.width="90%", out.height="130%", fig.asp=1, dpi=1000, fig.align='center'}

From eebf5d48c465c07cb56eaf413b7969dbfe53f81b Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 14 Aug 2023 15:18:47 +0200
Subject: [PATCH 105/124] prettier

---
 environment.yml | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/environment.yml b/environment.yml
index 57e1e3f1..cede896c 100644
--- a/environment.yml
+++ b/environment.yml
@@ -44,4 +44,4 @@ dependencies:
   - conda-forge::r-sessioninfo=1.2.2
   - conda-forge::r-svglite=2.1.1
   - conda-forge::r-tidyverse=2.0.0
-  - conda-forge::r-yaml=2.3.7
\ No newline at end of file
+  - conda-forge::r-yaml=2.3.7

From 2ba05785d01d8fc7aa3835cabd4d40009fe7825d Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 14 Aug 2023 15:52:20 +0200
Subject: [PATCH 106/124] elaborated further about batch effect in docs

---
 docs/usage.md | 1 +
 1 file changed, 1 insertion(+)

diff --git a/docs/usage.md b/docs/usage.md
index 2f6719e4..3645e2b3 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -351,6 +351,7 @@ Option needed to account for batch effects in the data. To control for batch eff
 
 Then the DESeq2 script calculates the contrasts as usual, the batch effect just needs to be considered during the design definition.
 For more information, please check the [DESeq2 vignette](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html).
+Please note: Setting the `--batch_effect` option will NOT remove such effects from your data. The parameter is only used to visualize the effects by generating two sets of PCA and boxplots instead of only one; one set of plots will be generated from the input data, the other from the results of the batch correction in order to illustrate how the batches affect data.
 
 ### `--min_DEG_pathway`
 

From 255b1527bb2939ccb1284cc356f21269c3ae7bb6 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 14 Aug 2023 16:08:55 +0200
Subject: [PATCH 107/124] Fixed md5sum and incorrect indents

---
 assets/RNAseq_report.Rmd | 44 ++++++++++++++++++++--------------------
 tests/test_smrnaseq.yml  |  2 +-
 2 files changed, 23 insertions(+), 23 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 05a416a3..4cf6f3d9 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -2261,28 +2261,28 @@ cat("No software versions file was provided for primary analysis.")
 # In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is clearer in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html
 
 if (params$input_type == "featurecounts"){
-    cat(paste0(
-    "## RNAseq data analysis
-
-    The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(software_versions_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
-    "` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
-    as.character(ifelse(software_versions_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
-    "` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
-    Subsequently, adapter trimming was conducted with `Trim Galore ",
-    as.character(ifelse(software_versions_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
-    "` [^4] [@krueger2012trim]. `STAR v",
-    as.character(ifelse(software_versions_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
-    "` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
-    The RNA-seq data quality control was performed with `RSeQC ",
-    as.character(ifelse(software_versions_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
-    "` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
-    quant_tool,
-    " ",
-    quant_version,
-    "` ",
-    quant_cite,
-    "."
-    ))
+cat(paste0(
+"## RNAseq data analysis
+
+The Nextflow-based nf-core pipeline `rnaseq ", as.character(ifelse(software_versions_csv, version["nf-core/rnaseq"], version["Workflow"][[1]]["nf-core/rnaseq"])),
+"` [^1] was used for the RNAseq Bioinfomatics analysis. `FASTQC ",
+as.character(ifelse(software_versions_csv, version["FastQC"], version["FASTQC"][[1]]["fastqc"])),
+"` [^3] [@andrews2010fastqc] was used to determine quality of the FASTQ files.
+Subsequently, adapter trimming was conducted with `Trim Galore ",
+as.character(ifelse(software_versions_csv, version["Trim Galore!"], version["TRIMGALORE"][[1]]["trimgalore"])),
+"` [^4] [@krueger2012trim]. `STAR v",
+as.character(ifelse(software_versions_csv, substring((version["STAR"]), 7, ), version["MAKE_TRANSCRIPTS_FASTA"][[1]]["star"])),
+"` [@Dobin2013] aligner was used to map the reads that passed the quality control to the reference genome.
+The RNA-seq data quality control was performed with `RSeQC ",
+as.character(ifelse(software_versions_csv, version["RSeQC"], version["RSEQC_BAMSTAT"][[1]]["rseqc"])),
+"` [@wang2012rseqc] and read quantification of the features (e.g. genes) with `",
+quant_tool,
+" ",
+quant_version,
+"` ",
+quant_cite,
+"."
+))
 } else if (params$input_type == "smrnaseq"){
 cat(paste0(
 "## smRNA-seq data analysis
diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml
index 506bf6cf..34ff104b 100644
--- a/tests/test_smrnaseq.yml
+++ b/tests/test_smrnaseq.yml
@@ -4,7 +4,7 @@
     - test_smrnaseq
   files:
     - path: results_test/differential_gene_expression/metadata/smrnaseq_files.txt
-      md5sum: ce3cc117fc953a8ff4d17d0118123faf
+      md5sum: 80efe9419bec686191344c6ce9a4ece7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_clone9_vs_clone1.tsv
       md5sum: 9526e0b2be54b49a8d1694b6cdd939b7
     - path: results_test/differential_gene_expression/DE_genes_tables/DE_contrast_condition_treatment_control_vs_clone1.tsv

From ffafaa8b31b01b076c74d7288a91ed77fd82184b Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 15 Aug 2023 11:24:24 +0200
Subject: [PATCH 108/124] added boxplots after batcheffect; boxplots now from
 transformed data

---
 CHANGELOG.md             |   3 +-
 assets/RNAseq_report.Rmd | 128 +++++++++++++++++++++++++++++++++------
 testdata/onecontrast.tsv |   4 --
 3 files changed, 110 insertions(+), 25 deletions(-)
 delete mode 100644 testdata/onecontrast.tsv

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 9d2edb00..1d25343b 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics. Heatmaps are now equally ordered in zip and report
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
@@ -17,6 +16,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Boxplots of genes are now generated from rlog/vst counts instead of raw counts. Also, if batch effect correction is enabled, boxplots will be generated before and after correction
+- [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics. Heatmaps are now equally ordered in zip and report
 - [#211](https://github.com/qbic-pipelines/rnadeseq/pull/21) Replaced heatmaply with pheatmap for static plots and removed kaleido and reticulate from container
 - [#209](https://github.com/qbic-pipelines/rnadeseq/pull/209) Template update to 2.9, Chromium Falcon; exchanged file.exists for nf-core validation checks; changed round_DE param to int
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_library_scaled_gene_counts.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 4cf6f3d9..dfd734eb 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1087,8 +1087,15 @@ ggsave(plot = pca_with_labels, filename = "differential_gene_expression/plots/PC
 # Create interactive plot for report
 pca <- ggplotly(pca, tooltip = "text", width=750, height=400)
 
-#add export-as-SVG button to plot
+# Add export-as-SVG button to plot
 config(pca, modeBarButtonsToAdd = list(svg_exp)) %>% layout()
+
+# Save non-corrected normalized object for boxplots
+if (run_rlog) {
+    rld_noncorrected <- rld
+} else {
+    vsd_noncorrected <- vsd
+}
 ```
 
 \
@@ -1101,8 +1108,10 @@ A PCA of the batch-effect corrected data is shown below and can be found [here](
 ```{r PCA_ifBatch_plot, echo=F, message=F, warning=F, dpi=1200, fig.align='center', eval=params$batch_effect}
 if (run_rlog) {
     assay(rld) <- limma::removeBatchEffect(assay(rld), rld$batch)
+    rld_corrected <- rld
 } else {
     assay(vsd) <- limma::removeBatchEffect(assay(vsd), vsd$batch)
+    vsd_corrected <- vsd
 }
 pcaData2 <- plotPCA(if (run_rlog) rld else vsd, intgroup=c("combfactor"), ntop = dim(if (run_rlog) rld else vsd)[1], returnData=TRUE)
 percentVar <- round(100*attr(pcaData, "percentVar"))
@@ -1569,7 +1578,13 @@ pg[['x']][['layout']][['annotations']][[1]][['y']] <- -params$adj_pval_threshold
 config(pg, modeBarButtonsToAdd = list(svg_exp)) %>% layout(margin = list(l=80, b = ifelse(length(allgenes_files)>2, 150, 100)), width=700, height = 500*length(allgenes_files))
 ```
 
-```{r box_plots, echo=F, message =F}
+```{r box_plots_noBatch, echo=F, message=F}
+# Decide which object to use for boxplots without batch correction
+if (run_rlog) {
+    box_object <- rld_noncorrected
+} else {
+    box_object <- vsd_noncorrected
+}
 
 ############### BOXPLOTS GENE EXPRESSION PER CONDITION ##########################
 # extract ID for genes to plot, make 20 plots:
@@ -1580,14 +1595,21 @@ if (length(DE_genes_plot) > 20) {
 } else {
     random_DE_genes_plot = DE_genes_plot
 }
-for (i in random_DE_genes_plot){
-    colData(cds)$combfactor_contrasts <- apply(as.data.frame(metadata_save[ ,conditions_contrasts]),1,paste, collapse = "_")
-    boxplot_counts <- plotCounts(cds, gene=i, intgroup=c("combfactor_contrasts"), returnData=TRUE, normalized = T)
-    boxplot_counts$variable = row.names(boxplot_counts)
-    plot <- ggplot(data=boxplot_counts, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
+
+colData(box_object)$combfactor_contrasts <- apply(as.data.frame(metadata_save[ ,conditions_contrasts]),1,paste, collapse = "_")
+
+# Previously, the function plotCounts was used here before ggplot. In PR #215, this was changed in order to allow for plotting the rlog/vst-normalized counts before/after batch correction (plotCounts can only do a normalization by sizeFactors). This was done following the instructions here: https://support.bioconductor.org/p/68859/
+# 1. Get columns from coldata
+box_df <-  data.frame(colData(box_object))
+
+for (i in random_DE_genes_plot) {
+
+    # 2. Get 1 column from assay; this column will correspond to the target gene
+    box_df$count <- assay(box_object)[i,]
+    plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
                 geom_boxplot(position=position_dodge()) +
                 geom_jitter(position=position_dodge(.8)) +
-                ggtitle(paste("Gene ",i,sep="")) + xlab("") + ylab("Normalized gene counts") + theme_bw() +
+                ggtitle(paste("Gene ",i,sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
                 theme(text = element_text(size=12),
                 axis.text.x = element_text(angle=45, vjust=1,hjust=1))
     ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,".svg",sep=""), width=10, height=5, plot=plot)
@@ -1596,7 +1618,7 @@ for (i in random_DE_genes_plot){
 }
 
 # make boxplots of interesting genes in gene list
-if (isProvided(params$path_genelist)){
+if (isProvided(params$path_genelist)) {
     gene_ids <- read.table(params$path_genelist, col.names = "requested_gene_name")
     write.table(gene_ids, file="differential_gene_expression/metadata/requested_gene_list.txt", col.names=F, row.names=F, sep="\t")
     gene_ids$requested_gene_name <- sapply(gene_ids$requested_gene_name, toupper)
@@ -1605,23 +1627,89 @@ if (isProvided(params$path_genelist)){
     # get Ensemble IDs from requested genes
     requested_genes_plot <- subset(gene_names, gene_name %in% gene_ids$requested_gene_name)
 
-    # Check that genes are in the cds table
-    requested_genes_plot <- subset(requested_genes_plot, requested_genes_plot$Ensembl_ID %in% row.names(cds))
+    # Check that genes are in the box_object table
+    requested_genes_plot <- subset(requested_genes_plot, requested_genes_plot$Ensembl_ID %in% row.names(box_object))
     requested_genes_plot_Ensembl <- requested_genes_plot$Ensembl_ID
     requested_genes_plot_gene_name <- requested_genes_plot$gene_name
+
     for (i in seq_along(requested_genes_plot_Ensembl)) {
-        boxplot_counts <- plotCounts(cds, gene=requested_genes_plot_Ensembl[i], intgroup=c("combfactor_contrasts"), returnData=TRUE, normalized = T)
-        boxplot_counts$variable = row.names(boxplot_counts)
-        plot <- ggplot(data=boxplot_counts, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
-        geom_boxplot(position=position_dodge()) +
-        geom_jitter(position=position_dodge(.8)) +
-        ggtitle(paste("Gene ",requested_genes_plot_gene_name[i],sep="")) + xlab("") + ylab("Normalized gene counts") + theme_bw() +
-        theme(text = element_text(size=12),
-                axis.text.x = element_text(angle=45, vjust=1,hjust=1))
+
+        # As above
+        box_df$count <- assay(box_object)[requested_genes_plot_Ensembl[i],]
+        plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
+                    geom_boxplot(position=position_dodge()) +
+                    geom_jitter(position=position_dodge(.8)) +
+                    ggtitle(paste("Gene ",requested_genes_plot_gene_name[i],sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                    theme(text = element_text(size=12),
+                    axis.text.x = element_text(angle=45, vjust=1,hjust=1))
         ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".svg",sep=""), width=10, height=5, plot=plot)
         ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".png",sep=""), width=10, height=5, plot=plot)
         ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".pdf",sep=""), width=10, height=5, plot=plot)
-    }
+    }    
+}
+```
+
+```{r box_plots_Batch, echo=F, message=F}
+# Decide which object to use for boxplots without batch correction
+if (run_rlog) {
+    box_object <- rld_corrected
+} else {
+    box_object <- vsd_corrected
+}
+
+############### BOXPLOTS GENE EXPRESSION PER CONDITION ##########################
+
+
+colData(box_object)$combfactor_contrasts <- apply(as.data.frame(metadata_save[ ,conditions_contrasts]),1,paste, collapse = "_")
+
+# Previously, the function plotCounts was used here before ggplot. In PR #215, this was changed in order to allow for plotting the rlog/vst-normalized counts before/after batch correction (plotCounts can only do a normalization by sizeFactors). This was done following the instructions here: https://support.bioconductor.org/p/68859/
+# 1. Get columns from coldata
+box_df <-  data.frame(colData(box_object))
+
+for (i in random_DE_genes_plot) {
+
+    # 2. Get 1 column from assay; this column will correspond to the target gene
+    box_df$count <- assay(box_object)[i,]
+    plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
+                geom_boxplot(position=position_dodge()) +
+                geom_jitter(position=position_dodge(.8)) +
+                ggtitle(paste("Gene ",i,sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                theme(text = element_text(size=12),
+                axis.text.x = element_text(angle=45, vjust=1,hjust=1))
+    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.svg",sep=""), width=10, height=5, plot=plot)
+    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.png",sep=""), width=10, height=5, plot=plot)
+    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.pdf",sep=""), width=10, height=5, plot=plot)
+}
+
+# make boxplots of interesting genes in gene list
+if (isProvided(params$path_genelist)) {
+    gene_ids <- read.table(params$path_genelist, col.names = "requested_gene_name")
+    write.table(gene_ids, file="differential_gene_expression/metadata/requested_gene_list.txt", col.names=F, row.names=F, sep="\t")
+    gene_ids$requested_gene_name <- sapply(gene_ids$requested_gene_name, toupper)
+    gene_names$gene_name <- sapply(gene_names$gene_name, toupper)
+
+    # get Ensemble IDs from requested genes
+    requested_genes_plot <- subset(gene_names, gene_name %in% gene_ids$requested_gene_name)
+
+    # Check that genes are in the box_object table
+    requested_genes_plot <- subset(requested_genes_plot, requested_genes_plot$Ensembl_ID %in% row.names(box_object))
+    requested_genes_plot_Ensembl <- requested_genes_plot$Ensembl_ID
+    requested_genes_plot_gene_name <- requested_genes_plot$gene_name
+
+    for (i in seq_along(requested_genes_plot_Ensembl)) {
+
+        # As above
+        box_df$count <- assay(box_object)[requested_genes_plot_Ensembl[i],]
+        plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
+                    geom_boxplot(position=position_dodge()) +
+                    geom_jitter(position=position_dodge(.8)) +
+                    ggtitle(paste("Gene ",requested_genes_plot_gene_name[i],sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                    theme(text = element_text(size=12),
+                    axis.text.x = element_text(angle=45, vjust=1,hjust=1))
+        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.svg",sep=""), width=10, height=5, plot=plot)
+        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.png",sep=""), width=10, height=5, plot=plot)
+        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.pdf",sep=""), width=10, height=5, plot=plot)
+    }    
 }
 ```
 
diff --git a/testdata/onecontrast.tsv b/testdata/onecontrast.tsv
deleted file mode 100644
index f9404753..00000000
--- a/testdata/onecontrast.tsv
+++ /dev/null
@@ -1,4 +0,0 @@
-treatment_treated_vs_control
-0
-1
-0

From fd4e3360b46949951eef0270c38f4042fea8c8d5 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Tue, 15 Aug 2023 13:11:19 +0200
Subject: [PATCH 109/124] added logical to the boxplots after batcheffect;
 removed quotes from the gene_list table that is written to metadata

---
 assets/RNAseq_report.Rmd | 4 ++--
 1 file changed, 2 insertions(+), 2 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index dfd734eb..d2e9c409 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1649,7 +1649,7 @@ if (isProvided(params$path_genelist)) {
 }
 ```
 
-```{r box_plots_Batch, echo=F, message=F}
+```{r box_plots_Batch, echo=F, message=F, eval=params$batch_effect}
 # Decide which object to use for boxplots without batch correction
 if (run_rlog) {
     box_object <- rld_corrected
@@ -1684,7 +1684,7 @@ for (i in random_DE_genes_plot) {
 # make boxplots of interesting genes in gene list
 if (isProvided(params$path_genelist)) {
     gene_ids <- read.table(params$path_genelist, col.names = "requested_gene_name")
-    write.table(gene_ids, file="differential_gene_expression/metadata/requested_gene_list.txt", col.names=F, row.names=F, sep="\t")
+    write.table(gene_ids, file="differential_gene_expression/metadata/requested_gene_list.txt", col.names=F, row.names=F, sep="\t", quote=F)
     gene_ids$requested_gene_name <- sapply(gene_ids$requested_gene_name, toupper)
     gene_names$gene_name <- sapply(gene_names$gene_name, toupper)
 

From 823169f49e3948268fa52469b5682bfe5e330807 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 16 Aug 2023 14:47:15 +0200
Subject: [PATCH 110/124] replaced missing gene_names in gtf with gene_ids

---
 CHANGELOG.md             | 1 +
 assets/RNAseq_report.Rmd | 3 +++
 2 files changed, 4 insertions(+)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 9d2edb00..42f25c5c 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -17,6 +17,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#216](https://github.com/qbic-pipelines/rnadeseq/pull/216) For rsem/salmon-imports, when a GTF is sometimes missing gene_names, these are now replaced by gene_ids instead
 - [#211](https://github.com/qbic-pipelines/rnadeseq/pull/21) Replaced heatmaply with pheatmap for static plots and removed kaleido and reticulate from container
 - [#209](https://github.com/qbic-pipelines/rnadeseq/pull/209) Template update to 2.9, Chromium Falcon; exchanged file.exists for nf-core validation checks; changed round_DE param to int
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Changed < and > to <= and => for logF/pval comparisons, renamed gene_counts_tables/deseq2_library_scaled_gene_counts.tsv to deseq2_library_scaled_gene_counts.tsv and added entry to the folder explanation; renamed param pval_threshold to adj_pval_threshold
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index 4cf6f3d9..570e0efd 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -539,6 +539,9 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) {
     tx2gene_gtf[] <- lapply(tx2gene_gtf, function(x) gsub("\\.\\d+", "", x))
     colnames(tx2gene_gtf) <- c("transcript_id", "gene_id") #, "TXID"
     gene_names <- gtf[c("gene_id", "gene_name")]
+
+    # Where no gene_name is available, use gene_id instead
+    gene_names$gene_name <- ifelse(is.na(gene_names$gene_name), gene_names$gene_id, gene_names$gene_name)
     colnames(gene_names) <- c("Ensembl_ID", "gene_name")
     gene_names <- distinct(gene_names)
     rownames(gene_names) <- gene_names[,1]

From 0b7a721e47bb16e7265c2bc048bdb5f4ae113c75 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 16 Aug 2023 15:36:38 +0200
Subject: [PATCH 111/124] Boxplots now also have their gene names in title and
 filename

---
 CHANGELOG.md             |  1 +
 assets/RNAseq_report.Rmd | 42 ++++++++++++++++++++++------------------
 2 files changed, 24 insertions(+), 19 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 1d25343b..c570998b 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
+- [#216](https://github.com/qbic-pipelines/rnadeseq/pull/216) Added gene_name to boxplot titles and filenames
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index d2e9c409..eafa6e0a 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1589,7 +1589,7 @@ if (run_rlog) {
 ############### BOXPLOTS GENE EXPRESSION PER CONDITION ##########################
 # extract ID for genes to plot, make 20 plots:
 DE_genes_plot <- subset(DE_genes_final_table, outcome == "DE")
-DE_genes_plot = unique(DE_genes_plot$Ensembl_ID)
+DE_genes_plot = unique(DE_genes_plot[c("Ensembl_ID", "gene_name")])
 if (length(DE_genes_plot) > 20) {
     random_DE_genes_plot = sample(DE_genes_plot,size = 2)
 } else {
@@ -1601,20 +1601,21 @@ colData(box_object)$combfactor_contrasts <- apply(as.data.frame(metadata_save[ ,
 # Previously, the function plotCounts was used here before ggplot. In PR #215, this was changed in order to allow for plotting the rlog/vst-normalized counts before/after batch correction (plotCounts can only do a normalization by sizeFactors). This was done following the instructions here: https://support.bioconductor.org/p/68859/
 # 1. Get columns from coldata
 box_df <-  data.frame(colData(box_object))
-
-for (i in random_DE_genes_plot) {
+for (i in nrow(random_DE_genes_plot)) {
+    ensid <- random_DE_genes_plot$Ensembl_ID[i]
+    genename <- random_DE_genes_plot$gene_name[i]
 
     # 2. Get 1 column from assay; this column will correspond to the target gene
-    box_df$count <- assay(box_object)[i,]
+    box_df$count <- assay(box_object)[ensid,]
     plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
                 geom_boxplot(position=position_dodge()) +
                 geom_jitter(position=position_dodge(.8)) +
-                ggtitle(paste("Gene ",i,sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                ggtitle(paste0("Gene ",genename,"/",ensid)) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
                 theme(text = element_text(size=12),
                 axis.text.x = element_text(angle=45, vjust=1,hjust=1))
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,".svg",sep=""), width=10, height=5, plot=plot)
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,".png",sep=""), width=10, height=5, plot=plot)
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,".pdf",sep=""), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,".svg"), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,".png"), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,".pdf"), width=10, height=5, plot=plot)
 }
 
 # make boxplots of interesting genes in gene list
@@ -1639,7 +1640,7 @@ if (isProvided(params$path_genelist)) {
         plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
                     geom_boxplot(position=position_dodge()) +
                     geom_jitter(position=position_dodge(.8)) +
-                    ggtitle(paste("Gene ",requested_genes_plot_gene_name[i],sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                    ggtitle(paste0("Gene ",requested_genes_plot_gene_name[i],"/",requested_genes_plot_Ensembl[i])) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
                     theme(text = element_text(size=12),
                     axis.text.x = element_text(angle=45, vjust=1,hjust=1))
         ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".svg",sep=""), width=10, height=5, plot=plot)
@@ -1666,19 +1667,22 @@ colData(box_object)$combfactor_contrasts <- apply(as.data.frame(metadata_save[ ,
 # 1. Get columns from coldata
 box_df <-  data.frame(colData(box_object))
 
-for (i in random_DE_genes_plot) {
+for (i in nrow(random_DE_genes_plot)) {
+    ensid <- random_DE_genes_plot$Ensembl_ID[i]
+    genename <- random_DE_genes_plot$gene_name[i]
+
 
     # 2. Get 1 column from assay; this column will correspond to the target gene
-    box_df$count <- assay(box_object)[i,]
+    box_df$count <- assay(box_object)[ensid,]
     plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
                 geom_boxplot(position=position_dodge()) +
                 geom_jitter(position=position_dodge(.8)) +
-                ggtitle(paste("Gene ",i,sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                ggtitle(paste0("Gene ",genename,"/",ensid)) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
                 theme(text = element_text(size=12),
                 axis.text.x = element_text(angle=45, vjust=1,hjust=1))
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.svg",sep=""), width=10, height=5, plot=plot)
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.png",sep=""), width=10, height=5, plot=plot)
-    ggsave(filename=paste("differential_gene_expression/plots/boxplots_example_genes/",i,"_after_batchcorrect.pdf",sep=""), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.svg"), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.png"), width=10, height=5, plot=plot)
+    ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.pdf"), width=10, height=5, plot=plot)
 }
 
 # make boxplots of interesting genes in gene list
@@ -1703,12 +1707,12 @@ if (isProvided(params$path_genelist)) {
         plot <- ggplot(data=box_df, aes(x=combfactor_contrasts, y=count, fill=combfactor_contrasts)) +
                     geom_boxplot(position=position_dodge()) +
                     geom_jitter(position=position_dodge(.8)) +
-                    ggtitle(paste("Gene ",requested_genes_plot_gene_name[i],sep="")) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
+                    ggtitle(paste0("Gene ",requested_genes_plot_gene_name[i],"/",requested_genes_plot_Ensembl[i])) + xlab("") + ylab(paste0(ifelse(run_rlog, "rlog", "vst"), "-normalized gene counts")) + theme_bw() +
                     theme(text = element_text(size=12),
                     axis.text.x = element_text(angle=45, vjust=1,hjust=1))
-        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.svg",sep=""), width=10, height=5, plot=plot)
-        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.png",sep=""), width=10, height=5, plot=plot)
-        ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.pdf",sep=""), width=10, height=5, plot=plot)
+        ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.svg"), width=10, height=5, plot=plot)
+        ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.png"), width=10, height=5, plot=plot)
+        ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.pdf"), width=10, height=5, plot=plot)
     }    
 }
 ```

From d1a01df45670c6378dbb8f64d2bbc53e4e702b80 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Wed, 16 Aug 2023 15:38:44 +0200
Subject: [PATCH 112/124] Corrected changelog

---
 CHANGELOG.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index c570998b..c0e1ce98 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#216](https://github.com/qbic-pipelines/rnadeseq/pull/216) Added gene_name to boxplot titles and filenames
+- [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Added gene_name to boxplot titles and filenames
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably

From cbf391525be9f6cef54bae106950700f72920dbd Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 17 Aug 2023 11:54:58 +0200
Subject: [PATCH 113/124] fixed missing labels in PCA_label plots

---
 assets/RNAseq_report.Rmd | 6 +++---
 1 file changed, 3 insertions(+), 3 deletions(-)

diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index eafa6e0a..879a294f 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1079,7 +1079,7 @@ ggsave(plot = pca, filename = "differential_gene_expression/plots/PCA_plot.png",
 
 # Save static plot with samplename labels
 pca_with_labels <- pca +
-    geom_label_repel(aes(label=name), force=2, force_pull=0, size=2.8, box.padding=0.15,show.legend  = F)
+    geom_label_repel(aes(label=name), force=2, force_pull=0, size=2.8, box.padding=0.15,show.legend  = F, max.overlaps=20)
 ggsave(plot = pca_with_labels, filename = "differential_gene_expression/plots/PCA_plot_with_labels.pdf", device = "pdf", dpi = 300)
 ggsave(plot = pca_with_labels, filename = "differential_gene_expression/plots/PCA_plot_with_labels.svg", device = "svg", dpi = 150)
 ggsave(plot = pca_with_labels, filename = "differential_gene_expression/plots/PCA_plot_with_labels.png", device = "png", dpi = 150)
@@ -1131,7 +1131,7 @@ ggsave(plot = pca2, filename = "differential_gene_expression/plots/PCA_batch_cor
 # Save static plot with samplename labels
 
 pca2_with_labels <- pca2 +
-    geom_label_repel(aes(label=name), force=2, force_pull=0, size=2.8, box.padding=0.15,show.legend  = F)
+    geom_label_repel(aes(label=name), force=2, force_pull=0, size=2.8, box.padding=0.15,show.legend  = F, max.overlaps=20)
 ggsave(plot = pca2_with_labels, filename = "differential_gene_expression/plots/PCA_batch_corrected_plot_with_labels.pdf", device = "pdf", dpi = 300)
 ggsave(plot = pca2_with_labels, filename = "differential_gene_expression/plots/PCA_batch_corrected_plot_with_labels.svg", device = "svg", dpi = 150)
 ggsave(plot = pca2_with_labels, filename = "differential_gene_expression/plots/PCA_batch_corrected_plot_with_labels.png", device = "png", dpi = 150)
@@ -1172,7 +1172,7 @@ dist_heatmap
 dev.off()
 
 # Recompute heatmap with heatmaply for interactive plot in report
-dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col=colors, grid_color = "grey", Rowv = sampleDistClust, Colv = "Rowv")
+dist_heatmap <- heatmaply(sampleDistMatrix, dendrogram = "both", col = colors, grid_color = "grey", Rowv = sampleDistClust, Colv = "Rowv")
 ```
 
 ```{r heatmap_show, echo=FALSE, message=FALSE, warning=FALSE, out.width="90%", out.height="130%", fig.asp=1, dpi=1000, fig.align='center'}

From 4957b1475c1860feea956f6bb2a1359507bde0f9 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 17 Aug 2023 11:56:11 +0200
Subject: [PATCH 114/124] updated changelog

---
 CHANGELOG.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index c0e1ce98..da6900ab 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Added
 
-- [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Added gene_name to boxplot titles and filenames
+- [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Added gene_name to boxplot titles and filenames; increased threshold before overlapping PCA labels are hidden
 - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support
 - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions
 - [#206](https://github.com/qbic-pipelines/rnadeseq/pull/206) Added logic to decide between rlog and vst, added tryCatch for heatmap saving because this only works unreliably

From 3f4ac364105c714d57c9d8f616ea45b9ebc32784 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 17 Aug 2023 15:39:29 +0200
Subject: [PATCH 115/124] Software version bumps for release

---
 .github/workflows/ci.yml | 10 ++++++----
 CHANGELOG.md             |  4 +++-
 Dockerfile               |  6 +++---
 environment.yml          |  2 +-
 modules/local/report.nf  |  2 +-
 nextflow.config          |  2 +-
 6 files changed, 15 insertions(+), 11 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index 3996d01d..b0fac9f3 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -40,13 +40,14 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 # Change this version before/after release
 
+      # Change the third version here before/after release
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull ghcr.io/qbic-pipelines/rnadeseq:dev
-          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
@@ -93,13 +94,14 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 # Change this version before/after release
 
+       # Change the third version here before/after release
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
           docker pull ghcr.io/qbic-pipelines/rnadeseq:dev
-          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev
+          docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2
 
       - name: Install Nextflow
         uses: nf-core/setup-nextflow@v1
diff --git a/CHANGELOG.md b/CHANGELOG.md
index 8a340a75..1715a0c1 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -3,7 +3,7 @@
 The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
 and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).
 
-## 2.2dev
+## 2.2 Avenue of Poplars
 
 ### Added
 
@@ -17,6 +17,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
+- [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Release 2.2 to master
+- [#217](https://github.com/qbic-pipelines/rnadeseq/pull/217) Preparing release 2.2 with version bumps
 - [#216](https://github.com/qbic-pipelines/rnadeseq/pull/216) For rsem/salmon-imports, when a GTF is sometimes missing gene_names, these are now replaced by gene_ids instead
 - [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Boxplots of genes are now generated from rlog/vst counts instead of raw counts. Also, if batch effect correction is enabled, boxplots will be generated before and after correction
 - [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics. Heatmaps are now equally ordered in zip and report
diff --git a/Dockerfile b/Dockerfile
index 4f8ce7fe..c13267b3 100644
--- a/Dockerfile
+++ b/Dockerfile
@@ -5,14 +5,14 @@ LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oska
 LABEL org.opencontainers.image.licenses=MIT
 COPY environment.yml /
 #RUN conda install -c conda-forge mamba
-RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \
+RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.2 && \
     mamba clean --all --yes
 RUN apt-get update -qq && \
     apt-get install -y zip procps ghostscript
 # Add conda installation dir to PATH
-ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH
+ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.2/bin:$PATH
 # Dump the details of the installed packates to a file for posterity
-RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml
+RUN mamba env export --name qbic-pipelines-rnadeseq-2.2 > qbic-pipelines-rnadeseq-2.2.yml
 # Instruct R processes to use these empty files instead of clashing with a local config
 RUN touch .Rprofile
 RUN touch .Renviron
diff --git a/environment.yml b/environment.yml
index cede896c..cf3dc64d 100644
--- a/environment.yml
+++ b/environment.yml
@@ -1,7 +1,7 @@
 # You can use this file to create a conda environment for this pipeline:
 #   conda env create -f environment.yml
 # use this to find packages: https://anaconda.org/
-name: qbic-pipelines-rnadeseq-dev
+name: qbic-pipelines-rnadeseq-2.2
 channels:
   - bioconda
   - conda-forge
diff --git a/modules/local/report.nf b/modules/local/report.nf
index 167fceb5..eac509d0 100644
--- a/modules/local/report.nf
+++ b/modules/local/report.nf
@@ -1,6 +1,6 @@
 process REPORT {
 
-    container 'ghcr.io/qbic-pipelines/rnadeseq:dev'
+    container 'ghcr.io/qbic-pipelines/rnadeseq:2.2'
 
     input:
     path gene_counts
diff --git a/nextflow.config b/nextflow.config
index 866a45e4..9018f037 100644
--- a/nextflow.config
+++ b/nextflow.config
@@ -252,7 +252,7 @@ manifest {
     description     = """Differential gene expression analysis and pathway analysis of RNAseq data"""
     mainScript      = 'main.nf'
     nextflowVersion = '!>=23.04.0'
-    version         = '2.2dev'
+    version         = '2.2'
     doi             = ''
 }
 

From 92db42dfe61d1a1b94168fcfc4338ad3a759bd45 Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Thu, 17 Aug 2023 15:42:58 +0200
Subject: [PATCH 116/124] modified some comments

---
 .github/workflows/ci.yml | 8 ++++----
 1 file changed, 4 insertions(+), 4 deletions(-)

diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml
index b0fac9f3..e40f2eb6 100644
--- a/.github/workflows/ci.yml
+++ b/.github/workflows/ci.yml
@@ -40,9 +40,9 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 # Change this version before/after release
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2
 
-      # Change the third version here before/after release
+      # Change the version above and the third version below before/after release
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |
@@ -94,9 +94,9 @@ jobs:
             environment.yml
       - name: Build new docker image
         if: env.MATCHED_FILES
-        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 # Change this version before/after release
+        run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2
 
-       # Change the third version here before/after release
+      # Change the version above and the third version below before/after release
       - name: Pull docker image
         if: ${{ !env.MATCHED_FILES }}
         run: |

From cf4487f981568b81aff3bf8e2707b15c9096ba5c Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 18 Aug 2023 09:30:06 +0200
Subject: [PATCH 117/124] Fixed changelog

---
 CHANGELOG.md | 6 +++---
 1 file changed, 3 insertions(+), 3 deletions(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index 1715a0c1..9643cd01 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -17,9 +17,9 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
-- [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Release 2.2 to master
-- [#217](https://github.com/qbic-pipelines/rnadeseq/pull/217) Preparing release 2.2 with version bumps
-- [#216](https://github.com/qbic-pipelines/rnadeseq/pull/216) For rsem/salmon-imports, when a GTF is sometimes missing gene_names, these are now replaced by gene_ids instead
+- [#219](https://github.com/qbic-pipelines/rnadeseq/pull/219) Release 2.2 to master
+- [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Preparing release 2.2 with version bumps
+- [#217](https://github.com/qbic-pipelines/rnadeseq/pull/217) For rsem/salmon-imports, when a GTF is sometimes missing gene_names, these are now replaced by gene_ids instead
 - [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Boxplots of genes are now generated from rlog/vst counts instead of raw counts. Also, if batch effect correction is enabled, boxplots will be generated before and after correction
 - [#214](https://github.com/qbic-pipelines/rnadeseq/pull/214) Added smrnaseq input support -->follow-up to check that the CI test works. Modified smrnaseq testdata to be like QBiC datasets. Updated container env and fixed a resulting bug with include_graphics. Heatmaps are now equally ordered in zip and report
 - [#211](https://github.com/qbic-pipelines/rnadeseq/pull/21) Replaced heatmaply with pheatmap for static plots and removed kaleido and reticulate from container

From 0f63439ca5b452d03d4ba2b390c54380b3c797f4 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Fri, 18 Aug 2023 15:52:28 +0200
Subject: [PATCH 118/124] Update nextflow_schema.json

Co-authored-by: Louisa Perelo <44900284+louperelo@users.noreply.github.com>
---
 nextflow_schema.json | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/nextflow_schema.json b/nextflow_schema.json
index e992d9cd..c3bc11d7 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -87,7 +87,7 @@
                     "type": "string",
                     "default": "rlog",
                     "enum": ["rlog", "vst", "vst-force"],
-                    "description": "Which transformation to use during DE analysis; either rlog or vst or vst-force . If vst: the pipeline will override this and run rlog if it detects a strong variation in size factors in the input data (this will be stated in the report). To disable this behavior, set to vst-force."
+                    "description": "Which transformation to use during DE analysis; either rlog or vst or vst-force. If vst: the pipeline will override this and run rlog if it detects a strong variation in size factors in the input data (this will be stated in the report). To disable this behavior, set to vst-force."
                 },
                 "vst_genes_number": {
                     "type": "integer",

From a83a94b9b0eb6e285e5d0a71ed2690981f220770 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Fri, 18 Aug 2023 15:56:06 +0200
Subject: [PATCH 119/124] Update nextflow_schema.json

Co-authored-by: Louisa Perelo <44900284+louperelo@users.noreply.github.com>
---
 nextflow_schema.json | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/nextflow_schema.json b/nextflow_schema.json
index c3bc11d7..8c39ea53 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -159,7 +159,7 @@
                 },
                 "quote": {
                     "type": "string",
-                    "description": "Quote file to link in the report‚."
+                    "description": "Quote file to link in the report."
                 },
                 "software_versions": {
                     "type": "string",

From ccb67df49d4eb7b1e140248232ab8cab9f5b6fc7 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Fri, 18 Aug 2023 15:57:25 +0200
Subject: [PATCH 120/124] Update nextflow_schema.json

Co-authored-by: Louisa Perelo <44900284+louperelo@users.noreply.github.com>
---
 nextflow_schema.json | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/nextflow_schema.json b/nextflow_schema.json
index 8c39ea53..93814eb8 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -339,7 +339,7 @@
             "type": "object",
             "fa_icon": "fab fa-acquisitions-incorporated",
             "description": "Set the top limit for requested resources for any single job.",
-            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
+            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system. \n Note that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
             "properties": {
                 "max_cpus": {
                     "type": "integer",

From 28d8763f4d26133d01a9d54e6c940f20448f48f5 Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Fri, 18 Aug 2023 15:57:58 +0200
Subject: [PATCH 121/124] Update docs/usage.md

Co-authored-by: Louisa Perelo <44900284+louperelo@users.noreply.github.com>
---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index 3645e2b3..ebc04068 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -538,7 +538,7 @@ The pipeline also dynamically loads configurations from [https://github.com/nf-c
 Note that multiple profiles can be loaded, for example: `-profile test,docker` - the order of arguments is important!
 They are loaded in sequence, so later profiles can overwrite earlier profiles.
 
-If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended, since it can lead to different results on different machines dependent on the computer enviroment.
+If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended, since it can lead to different results on different machines dependent on the computer environment.
 
 - `test`
   - A profile with a complete configuration for automated testing

From 1860521ed0d129b43cf17a2f75b67ec269c288ca Mon Sep 17 00:00:00 2001
From: WackerO <43847497+WackerO@users.noreply.github.com>
Date: Fri, 18 Aug 2023 15:58:16 +0200
Subject: [PATCH 122/124] Update docs/usage.md

Co-authored-by: Louisa Perelo <44900284+louperelo@users.noreply.github.com>
---
 docs/usage.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/docs/usage.md b/docs/usage.md
index ebc04068..2d74b416 100644
--- a/docs/usage.md
+++ b/docs/usage.md
@@ -542,7 +542,7 @@ If `-profile` is not specified, the pipeline will run locally and expect all sof
 
 - `test`
   - A profile with a complete configuration for automated testing
-  - Includes links to test data so needs no other parameters
+  - Includes links to test data, therefore needs no other parameters
 - `docker`
   - A generic configuration profile to be used with [Docker](https://docker.com/)
 - `singularity`

From 4739c0a0ed73786d5b18810e953cb3824b246e5e Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Fri, 18 Aug 2023 15:59:28 +0200
Subject: [PATCH 123/124] smaller clean-up changes

---
 .github/ISSUE_TEMPLATE/config.yml |  7 -------
 assets/RNAseq_report.Rmd          | 13 +++++++------
 nextflow_schema.json              |  2 +-
 3 files changed, 8 insertions(+), 14 deletions(-)
 delete mode 100644 .github/ISSUE_TEMPLATE/config.yml

diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml
deleted file mode 100644
index a5de823c..00000000
--- a/.github/ISSUE_TEMPLATE/config.yml
+++ /dev/null
@@ -1,7 +0,0 @@
-contact_links:
-  - name: Join nf-core
-    url: https://nf-co.re/join
-    about: Please join the nf-core community here
-  - name: "Slack #rnadeseq channel"
-    url: https://nfcore.slack.com/channels/rnadeseq
-    about: Discussion about the qbic-pipelines/rnadeseq pipeline
diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd
index bd3aac6d..b1aa99b8 100644
--- a/assets/RNAseq_report.Rmd
+++ b/assets/RNAseq_report.Rmd
@@ -1,4 +1,5 @@
 ---
+# Written by Gisela Gabernet, Stefan Czemmel, Silvia Morini, Oskar Wacker, QBiC and released under the MIT License
 output:
     html_document:
         toc: true # table of contents
@@ -671,7 +672,7 @@ if (params$norm_method == "vst-force") {
     norm_method_long_text <- "variance stabilizing transformation (vst)"
     nsub_genes_text <- paste0("The number of subset genes for the vst transformation was set to ", as.character(params$nsub_genes), ".")
     if (var_sizeFactors >= vst_cutoff) {
-        norm_method_override_text <- paste0(" This was due to using the `vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
+        norm_method_override_text <- paste0(" This was due to using the `--norm_method vst-force` argument. It is recommended to rerun the pipeline using the rlog transformation as the size factors of the different samples have a variance of ~", round(var_sizeFactors, 4), ". For high sample variances (i.e. different sequencing depths between samples), the rlog transformation is more suitable [@love2014differential].")
         nsub_genes_text <- ""
     }
 } else if (params$norm_method == "vst") {
@@ -945,7 +946,7 @@ The read mapping statistics to the reference genome for each sample are shown be
 ```
 
 ```{r STAR_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1800, fig.cap="STAR: Mapping Statistics", fig.align='center'}
-# Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every call to not fail with some stupid error
+# Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every include_graphic() call to not fail with an error
 knitr.graphics.rel_path = F
 knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"), error=F)
 ```
@@ -1415,7 +1416,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_
         write.table(DE_genes_contrast_genename, file=paste("differential_gene_expression/allgenes/DE_contrast_",contname,".tsv",sep=""), sep="\t", quote=F, col.names = T, row.names = F)
 
         # Select only significantly DE
-        DE_genes_contrast <-  subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
+        DE_genes_contrast <- subset(DE_genes_contrast_genename, padj <= params$adj_pval_threshold & abs(log2FoldChange) >= params$logFC_threshold)
         DE_genes_contrast <- DE_genes_contrast[order(DE_genes_contrast$padj),]
         # Round table before writing
         if (isProvided(params$round_DE) && params$round_DE>-1) {
@@ -1574,8 +1575,8 @@ pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_
         scale_x_continuous(breaks = seq(-200, 200, ifelse(log2FoldChange_quotient<3, 1, 5)))
 
 pg <- ggplotly(pg, tooltip = "text")
-pg[['x']][['layout']][['annotations']][[2]][['x']] <- -params$adj_pval_threshold #-0.05 TODO: what do these two lines do?
-pg[['x']][['layout']][['annotations']][[1]][['y']] <- -params$adj_pval_threshold
+pg[['x']][['layout']][['annotations']][[2]][['x']] <- -0.05 # These lines add a bit of additional space between the x/y axis labels and the plot
+pg[['x']][['layout']][['annotations']][[1]][['y']] <- -0.05
 
 #add export button to plot
 config(pg, modeBarButtonsToAdd = list(svg_exp)) %>% layout(margin = list(l=80, b = ifelse(length(allgenes_files)>2, 150, 100)), width=700, height = 500*length(allgenes_files))
@@ -2326,7 +2327,7 @@ if (software_versions_csv) {
         quant_version <- as.character(version["featureCounts"])
         quant_cite <- "[@liao2014featurecounts]"
     }
-} else if (!(software_versions_csv)) {
+} else {
     if (params$input_type == "rsem") {
         quant_tool <- "RSEM"
         quant_version <- as.character(version["RSEM_CALCULATEEXPRESSION"][[1]]["rsem"])
diff --git a/nextflow_schema.json b/nextflow_schema.json
index e992d9cd..7b7ebc8c 100644
--- a/nextflow_schema.json
+++ b/nextflow_schema.json
@@ -339,7 +339,7 @@
             "type": "object",
             "fa_icon": "fab fa-acquisitions-incorporated",
             "description": "Set the top limit for requested resources for any single job.",
-            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
+            "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.",
             "properties": {
                 "max_cpus": {
                     "type": "integer",

From 7c33a84f51eb1857f1ae589baae25d2ecc196e9a Mon Sep 17 00:00:00 2001
From: WackerO <oskar.wacker@web.de>
Date: Mon, 21 Aug 2023 08:28:45 +0200
Subject: [PATCH 124/124] modified changelog

---
 CHANGELOG.md | 2 +-
 1 file changed, 1 insertion(+), 1 deletion(-)

diff --git a/CHANGELOG.md b/CHANGELOG.md
index be720dd5..56d1d982 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -17,7 +17,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ### Changed
 
-- [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Release 2.2 to master
+- [#219](https://github.com/qbic-pipelines/rnadeseq/pull/219) Release 2.2 to master
 - [#220](https://github.com/qbic-pipelines/rnadeseq/pull/220) Commit suggestions from PR review for release 2.2
 - [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Preparing release 2.2 with version bumps
 - [#217](https://github.com/qbic-pipelines/rnadeseq/pull/217) For rsem/salmon-imports, when a GTF is sometimes missing gene_names, these are now replaced by gene_ids instead