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Today’s goal:

In this session, we will use the feature counts table generated by the star_salmon job to run a statistical analysis to identify differentially expressed genes.

Log into the HPC

ssh userID@lyra.qut.edu.au

To go to a shared directory for your project, for example: “kenna_team”. Type the following command and hit enter:

cd /work/kenna_team/

Go to the results of the previous Session 3:

cd jess/session3/

Changing the folder permission for other members in the group to access the ‘results’ and ‘work’ folders:

chmod -R g+rwX results

chmod -R g+rwX work

Go to the folder with the feature counts generated by star_salmon:

cd run1_star_salmon/results/star_salmon

Various ‘salmon.merged.*’ files are generated for all the samples processed in the experiment. Let’s look for the file that contains the feature counts for genes:

head -25 salmon.merged.gene_counts.tsv

To browse the whole file type the following:

less salmon.merged.gene_counts.tsv

Copy the “salmon.merged.gene_counts.tsv” to your laptop and we will use it for statistical analysis using BioJupies (see below)

Differential gene expression analysis

Differential expression analysis using https://maayanlab.cloud/biojupies/analyze

From session 2, we will use the feature counts generated using the ‘star_salmon’ parameter setting. Either copy the relevant files (see below) to your laptop from the HPC or download the files below.

File

File name

File

Gene feature counts

salmon_merged.gene_counts.tsv

 

Transcript feature counts

salmon_merged.transcript_counts.tsv

 

Reading:

https://www.nature.com/articles/srep25533

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