#!/bin/bash # This script should be run on your local laptop or computer # Please make sure you have installed Docker Desktop using the instructions at: https://www.docker.com/products/docker-desktop/ # Before running this script, do the following ## 1. open the Docker Dekstop and make sure it is running ## 2. open a new terminal (MacOS) or command prompt (Windows) window #check if docker is running #the below command will print out the docker version if docker is running docker --version #get the docker image from https://hub.docker.com/r/nfcore/rnaseq/ #we will be using this image for all the analyses docker pull nfcore/rnaseq #check if the docker image was downloaded successfully docker images #spin up a docker container ##for M1 chip mac, use the below command docker run --platform linux/amd64 --name rna_bash --rm -it -v ~/Downloads/Intro_to_RNA-seq_data_analysis:/home nfcore/rnaseq bash ##for all other computers, use the below command docker run --name rna_bash --rm -it -v ~/Downloads/Intro_to_RNA-seq_data_analysis:/home nfcore/rnaseq bash #a new prompt will apear and the conatiner is now active #the home directory should have all the contents of ~/Downloads/Intro_to_RNA-seq_data_analysis #go to the home directory in the docker container cd home #run fastqc fastqc Bacteria_GATTACA_L001_R1_001.fastq #trim the reads using cutadapt cutadapt -a file:Adapter_Sequence.fasta -o trimmed.fastq Bacteria_GATTACA_L001_R1_001.fastq #run fastqc on the trimmed reads fastqc trimmed.fastq #create a new folder mkdir star_index #create the STAR index STAR --runMode genomeGenerate --genomeDir ./star_index --genomeFastaFiles rDNA_sequence.fasta --genomeSAindexNbases 3 #run STAR for the trimmed reads STAR --genomeDir ./star_index --readFilesIn ./trimmed.fastq #generate the read count matrix using featureCounts featureCounts -a rDNA.gtf -t CDS -o counts.txt Aligned.out.sam