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snakemake pipeline for mapping and pre-processing ALS RNA-seq

Scott Adamson
David Knowles / Tuuli Lappalainen labs

Installation:

First install this snakemake pipeline:

git clone git@github.com:scottiadamson/ALS_RNA-seq_snakemake_workflow.git
cd ALS_RNA-seq_snakemake_workflow

Then create a mamba environment with dependencies:

mamba create -n ALS_analysis_env
mamba activate ALS_analysis_env
mamba install --file requirements_updated.txt
pip install rnaseqc

Preparing references

Download the reference files if you haven't already:

cd references
wget https://ftp.ebi.ac.uk/pub/databases/gencode/Gencode_human/release_43/gencode.v43.transcripts.fa.gz
gunzip gencode.v43.transcripts.fa.gz
wget https://hgdownload.soe.ucsc.edu/goldenPath/hg38/bigZips/hg38.fa.gz
gunzip hg38.fa.gz
wget https://ftp.ebi.ac.uk/pub/databases/gencode/Gencode_human/release_43/gencode.v43.basic.annotation.gff3.gz
gunzip gencode.v43.basic.annotation.gff3.gz

Submit all of the reference building stuff
Be sure to edit the paths in the .sh files to the reference folder in your snakemake directory

sbatch kallisto_index.sh
sbatch labrat_index.sh
sbatch star_index.sh

Adjust config/config.yaml

Edit the config.yaml file to have the paths to this snakemake file, as well as the samples of interest.

Then run it:

cd ../ # only if you're still in the references folder
sbatch snakemake_submit.sh

Figuring out what's going on

If you just want to see the steps that the snakemake pipeline is running:

snakemake --snakefile Snakefile --dry-run -p -k