From edaacb7d0d7eadce7676b075dedfad8a03d3a5ed Mon Sep 17 00:00:00 2001 From: Leighton Pritchard Date: Mon, 27 May 2024 14:13:50 +0100 Subject: [PATCH] add a guide to template, conda, ncbi-genome-downlaod --- _quarto.yml | 2 + assets/images/primer_01.png | Bin 0 -> 126047 bytes assets/images/primer_env.jpg | Bin 0 -> 228341 bytes primer_env.qmd | 291 +++++++++++++++++++++++++++++++++++ 4 files changed, 293 insertions(+) create mode 100644 assets/images/primer_01.png create mode 100644 assets/images/primer_env.jpg create mode 100644 primer_env.qmd diff --git a/_quarto.yml b/_quarto.yml index 839b44b..203dfad 100644 --- a/_quarto.yml +++ b/_quarto.yml @@ -26,6 +26,8 @@ website: href: expectations.qmd - text: "Project and Time Management" href: time_management.qmd + - text: "Primer: Environment Setup" + href: primer_env.qmd - text: "Computational Biology" menu: - text: "Learning Resources" diff --git a/assets/images/primer_01.png b/assets/images/primer_01.png new file mode 100644 index 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zk*6|Ys}zhxKrlnm5DN;qTOlLr|3Lr-(uW3TF{U6fz!XdUd=>`jG+Wt#$Do568pU?0 z0|KnXgI!>XAfs3U1#lBm5VZM%VvIb(yjTG63;#*WJD-)rCL*Na*jFUEP@UsSp_|}= zq8)G|);hR~UD7@rCKV-CAM7+sVvCgtvU=y?Td{>QW^l@*G|kBYzYL%mc1byjN1S*Z zS-?pY6O%;4@DC~*st8fK0+}drH+bH8ma14|OuQ5f4vD>+NdTg1#BuEh?_O8?uEX&fd&c?6V6I88-L=L_keTdssYQ74rG@ct%Z#vpQp zD^45X(yw-PZY#aBcMXFG(Xk~3WK zQYH?dUtUyt literal 0 HcmV?d00001 diff --git a/primer_env.qmd b/primer_env.qmd new file mode 100644 index 0000000..50ad310 --- /dev/null +++ b/primer_env.qmd @@ -0,0 +1,291 @@ +--- +title: "Primer: Environment Setup" +image: ./assets/images/primer_env.jpg +description: | + Setting up your computational environment for the {{< var ay >}} project +number-sections: true +about: + template: marquee + links: + - icon: twitter + text: Twitter + href: https://twitter.com/scompbiol + - icon: github + text: Github + href: https://github.com/sipbs-compbiol + - icon: envelope + text: Email + href: mailto:leighton.pritchard@strath.ac.uk +html: + anchor-sections: true +--- + +This page runs through the process of setting up a computing environment similar to that you will need for your {{< var ay >}} project. + +::: { .callout-important } +## Prerequisites + +This primer assumes the following: + +- **you are working on your own computer** (e.g. a laptop/desktop machine) +- **you are able to start a terminal window** (e.g. `Terminal` on a Mac or Linux machine, or in Windows Subsystem Linux) + +You should either **be familiar with the command-line or have worked through the Software Carpentries shell lesson**: + +- [SWC Shell lesson](https://swcarpentry.github.io/shell-novice/) +::: + +The primer walks through the following steps: + +1. Downloading the SIPBSCompBiol project template repository +2. Creating and activating a `conda` environment for your project +3. Installing the `ncbi-genome-download` package and using it to download a set of genomes + +## Set up a project folder from a template + +::: { .callout-note } +Please read the Noble (2009) and Sandve _et al._ (2013) papers at the Ten Great Papers page, and the `README.md` file at the [template repository]() to understand the principles behind the organisation of this template. + +- [Ten Great Papers](https://widdowquinn.github.io/ten_great_papers/) +- [Bioinformatics project template](https://sipbs-compbiol.github.io/template_bioinformatics_project/) +- [Noble (2009)](http://doi.org/10.1371/journal.pcbi.1000424) +- [Sandve _et al._ (2013)](http://doi.org/10.1371/journal.pcbi.1003285) +::: + +### Download and extract the project template + +1. Use your browser to visit the project template repository at [https://github.com/sipbs-compbiol/template_bioinformatics_project](https://github.com/sipbs-compbiol/template_bioinformatics_project). +2. Click on the small triangle at the right of the green `Code` button to see the download options for the template. + +![Context menu for the `Code` download button](assets/images/primer_01.png) + +3. Click on `Download ZIP` to download a `.zip` format compressed file containing the repository template to your own computer, in the usual way you would download a file using your browser. This will place a file called `template_bioinformatics_project-master.zip` into that location. + +::: { .callout-tip collapse="true" } +## Where should I save this file? + +When you uncompress the repository template, it will expand into the full set of folders and subfolders. You can move this folder tree _after_ you have uncompressed the file, or you can move the `.zip` file to the directory where you want to keep the project files (e.g. under `Documents` on a Mac), and uncompress it there. The choice is yours. +::: + +4. Uncompress the file (e.g. by double-clicking on a Mac). This will produce a folder called `template_bioinformatics_project-master` in the same folder as the compressed file. + +::: { .callout-warning } +## Make sure you acturally uncompress the file + +Some operating systems, such as Windows, allow you to work with compressed files _as if they had been uncompressed_ without _actually_ uncompressing them. + +**You do need to fully uncompress the repository template to work with it.** +::: + +5. Start a terminal and navigate to the top-level folder in the repository template. + +::: { .callout-tip } +I saved the `.zip` file in the `Documents` folder on my computer, and uncompressed that file in the same location. My project template is therefore in the `~/Documents/template_bioinformatics_project-master` directory. I can navigate to this location using the `cd` command, and check the contents using the command `ls`. + +[**NOTE**: The `%` symbol is the command prompt, _not_ part of the command itself. This may look different on your computer (e.g. it might be a dollar sign: `$`).] + +```{bash} +% cd ~/Documents/template_bioinformatics_project-master +% ls +LICENSE README.md _config.yml assets/ data/ docs/ notebooks/ results/ scripts/ +``` +::: + +## Create and activate a `conda` environment for your project + +::: { .callout-warning } +Before beginning this part of the guide, you should make yourself familiar with the `conda` documentation at the link below: + +- [`conda` tutorial](https://docs.conda.io/projects/conda/en/latest/user-guide/getting-started.html) +::: + +::: { .callout-important } +## You will need to have already installed the `conda` package for your operating system. + +On Mac and Linux, this is usually a straightforward software installation, and you can follow the instructions at the link below: + +- [Installing Anaconda on Mac and Linux](https://docs.anaconda.com/free/anaconda/install/index.html) + +On Windows, you _must_ distinguish between installing Anaconda under Windows, and installing it under WSL (Windows Subsystem Linux). You will be using WSL for your project, and you **must** install `conda` within WSL (WSL _cannot_ see the version of `conda` installed under the Windows operating system; in any case the software that runs on a Linux OS will not typically run on a Windows OS). There are several online guides to help with this. + +- [Installing Conda/Anaconda in WSL](https://www.how2shout.com/how-to/install-anaconda-wsl-windows-10-ubuntu-linux-app.html) +- [Installing Miniconda in WSL](https://kontext.tech/article/1064/install-miniconda-and-anaconda-on-wsl-2-or-linux) +::: + +1. Create a new `conda` environment by issuing the command below. + +```{bash} +% conda create --name bm954_project -y +``` + +::: { .callout-note collapse="true" } +## Breaking down the components of the command line + +As above, the `%` symbol is the _command prompt_ - not part of the command itself. You do not type this when entering and running your command. + +- `conda`: you are running the `conda` program and instructing it to carry out an action +- `create`: this is the action you are instructing `conda` to undertake, to _create_ a new environment +- `--name bm954_project`: this _option_ tells `conda` what name the new environment is to be known by; you can choose any name you like, and do not have to use `bm954_project` if you would prefer to use something else. However, it is usually a good idea to name the environment such that you can tell immediately what it is used for +- `-y`: this answers all the optional questions about installation which `conda` might ask with "_yes_" +::: + +2. Activate the new `conda` environment you have just created by issuing the command below (if you called your environment something other than `bm954_project`, then use that name instead, here). + +```{bash} +% conda activate bm954_project +``` + +You should notice that the left-most part of your command prompt changes from `(base)` to `(bm954_project)` (or whatever name you used for your environment). + +::: { .callout-tip collapse="true"} +## How to check what environments are available + +You can use the command `conda info --envs` (demonstrated below) to list all the environments that your installation of `conda` is aware of: + +```{bash} +% conda info --envs +# conda environments: +# +base /Users/lpritc/opt/anaconda3 +2022_david /Users/lpritc/opt/anaconda3/envs/2022_david +2022_nora /Users/lpritc/opt/anaconda3/envs/2022_nora +algo_2022_py39 /Users/lpritc/opt/anaconda3/envs/algo_2022_py39 +aoc2021 /Users/lpritc/opt/anaconda3/envs/aoc2021 +aoc2022 /Users/lpritc/opt/anaconda3/envs/aoc2022 +aoc2023 /Users/lpritc/opt/anaconda3/envs/aoc2023 +bm432_py310 /Users/lpritc/opt/anaconda3/envs/bm432_py310 +bm954_project * /Users/lpritc/opt/anaconda3/envs/bm954_proj +[...] +``` +::: + +## Install `ncbi-genome-download` and recover some genomes + +One of the main advantages of `conda` is that it handles technical issues of _package management_ (the process of installing and configuring software tools) and makes using them much easier. The typical command we might use would be `conda install ` where `` is replaced by the name of the software you want to install. + +In particular, `conda` is a common software distribution route for bioinformatics software, through the `conda` _channel_ called [`bioconda`](https://bioconda.github.io/). + +::: { .callout-important } +In order to use `bioconda` on your computer, you will need to configure the channel. +::: + +### Configuring the `bioconda` channel + +::: { .callout-warning } +Before installing `bioconda` you should be awware of the information at the link below: + +- [`bioconda` tutorial](https://bioconda.github.io/tutorials/index.html) +::: + +To set up the `bioconda` channel on your computer, you should follow the instructions at the [`bioconda` website](https://bioconda.github.io/), and issue the four commands below, in sequence: + +```{bash} +conda config --add channels defaults +conda config --add channels bioconda +conda config --add channels conda-forge +conda config --set channel_priority strict +``` + +You only need to do this once on your computer, and `bioconda` will then be available in any environment you create. + +### Installing `ncbi-genome-download` + +To install the `ncbi-genome-download` package, we use the `conda install` command, as noted above. This will take a short while to download and configure all the _dependencies_ of the tool (packages that need to be installed for the software to run). The process you see in the terminal should resemble that shown below. + +```{bash} + % conda install ncbi-genome-download -y + Channels: + - conda-forge + - bioconda + - defaults +Platform: osx-64 +Collecting package metadata (repodata.json): +[...] +The following NEW packages will be INSTALLED: + + appdirs conda-forge/noarch::appdirs-1.4.4-pyh9f0ad1d_0 +[...] + xz conda-forge/osx-64::xz-5.2.6-h775f41a_0 + + + +Downloading and Extracting Packages: + +Preparing transaction: done +Verifying transaction: done +Executing transaction: done +``` + +Once this is complete, the `ncbi-genome-download` program should be available to you. You can test this by asking the software for its help guide, using the command `ncbi-genome-download -h` as demonstrated below: + +```{bash} +% ncbi-genome-download -h +usage: ncbi-genome-download [-h] [-s {refseq,genbank}] [-F FILE_FORMATS] [-l ASSEMBLY_LEVELS] [-g GENERA] [--genus GENERA] + [--fuzzy-genus] [-S STRAINS] [-T SPECIES_TAXIDS] [-t TAXIDS] [-A ASSEMBLY_ACCESSIONS] + [--fuzzy-accessions] [-R REFSEQ_CATEGORIES] [--refseq-category REFSEQ_CATEGORIES] [-o OUTPUT] + [--flat-output] [-H] [-P] [-u URI] [-p N] [-r N] [-m METADATA_TABLE] [-n] [-N] [-v] [-d] [-V] + [-M TYPE_MATERIALS] + groups +[...] +``` + +### Downloading a set of genome sequences + +::: { .callout-warning } +Before downloading genomes, you should familiarise yourself with the information at the links below: + +- [NCBI Assembly Level definitions](https://www.ncbi.nlm.nih.gov/datasets/docs/v2/glossary/#assembly-level) +- [`ncbi-genome-download` documentation](https://github.com/kblin/ncbi-genome-download/blob/master/README.md) +- [NCBI genome file formats](https://www.ncbi.nlm.nih.gov/datasets/docs/v2/reference-docs/file-formats/) +::: + +For your project, you will want to download all complete genome sequences for a single genus of bacteria. This section of the guide will illustrate that process for the genus _Dickeya_. + +The command to download all genomes of a single named genus of bacteria is `ncbi-genome-download --genera "" bacteria`, where `` is replaced by the actual name of the genus you want to download. + +To restrict downloaded genomes only to those that are _complete_, we use the option `--assembly-levels complete`. + +To download all available sequence data, including GBFF (annotated) and FASTA (sequence data only) we need to use the option `--formats all` + +If we wanted the program to keep us informed about what it was doing, we would ask it to be _verbose_ by using the option `--verbose` or `-v` + +Putting this together to download genomes for the genus _Dickeya_ we would use the command as demonstrated below: + +```{bash} +% ncbi-genome-download --assembly-levels complete --genera "Dickeya" --formats all --verbose bacteria +``` + +This will place the downloaded genomes into individual subdirectories, under the subdirectory `refseq`. These genome are compressed using the `gzip` program. We can tell this because the filnames end in `.gz`. + +```{bash} + % tree refseq +refseq +└── bacteria + ├── GCF_000147055.1 + │   ├── GCF_000147055.1_ASM14705v1_assembly_report.txt + │   ├── GCF_000147055.1_ASM14705v1_assembly_stats.txt + │   ├── GCF_000147055.1_ASM14705v1_cds_from_genomic.fna.gz + │   ├── GCF_000147055.1_ASM14705v1_feature_table.txt.gz + │   ├── GCF_000147055.1_ASM14705v1_genomic.fna.gz + │   ├── GCF_000147055.1_ASM14705v1_genomic.gbff.gz + │   ├── GCF_000147055.1_ASM14705v1_genomic.gff.gz + │   ├── GCF_000147055.1_ASM14705v1_protein.faa.gz + │   ├── GCF_000147055.1_ASM14705v1_protein.gpff.gz + │   ├── GCF_000147055.1_ASM14705v1_rna_from_genomic.fna.gz + │   ├── GCF_000147055.1_ASM14705v1_translated_cds.faa.gz +[...] +``` + +The downloaded files are in a number of different formats, and you will not need to use all of them in your project. + +## Summary + +This page provided a gude to setting up your project analogously to preparing your bench for laboratory work. + +You first used a template to set up your project's folder structure, which is a little like making sure you have a well-organised work area. + +You then installed `conda` and set up a new environment for your project, which plays a similar role to making your your bench is clean and practising a kind of "aseptic technique" for making sure that installed bioinformatics tools don't conflict with each other. + +You then installed `ncbi-genome-download` and obtained a set of bacterial genomes directly from NCBI. This is similar to acquiring a set of strains from a collection, and having them ready to work with on your benchtop. + +Your next steps using these genomes will be to prepare them so that they can be used in your experiments. \ No newline at end of file