This page provides a basic introduction to Unix commands to HPC users with no previous knowledge.
Log into HPC
ssh userID@lyra.qut.edu.au
Brief basic Unix commands recap
Once you log into the HPC, you will land in your personal home space (i.e. /home/myStudentID/). This space is only accessible to you. To work in collaboration with others we use workspaces (i.e. /work/myProjectName/).
To go to a shared directory for your project named “kenna_team” type the following command and hit enter:
cd /work/kenna_team/
Display list of files in a directory
ls -lh
Print working directory
pwd
Create a folder
mkdir myfolder
Enter new folder
cd myfolder
Move back to the previous folder
cd ..
Make a backup copy of the file
cp myfile.txt > myfile_copy.txt
Move a copy of a file to a newly created folder - note it is recommended to make a copy of important files prior to modifying or executing commands on them.
mv myfile_copy.txt myfolder/
View the content of a file (note hashtags # at the start of a line is used to provide information of the code underneath it)
#hash tags are used to add comments on what a command line does
#several commands can be used including cat, less, more, head and tail
cat myfile_copy.txt
#example: less -S allows to visualise very large (wide) files
less -S myfile_copy.txt
#stop viewing a file using the above command
--> Type “Control” and “c” at the same time.
Or “Control” and “d” at the same time.
#print the first 50 lines of a file
head -50 myfile_copy.txt
#print the last 20 lines of a file
tail -20 myfile_coy.txt
Go back to my personal space. Type 'cd' and hit enter. This will move you to /home/mystudentID/
cd
Running the nextflow nf-core/rnaseq pipeline
Requirements:
index.csv → a file that provides a list of sample IDs and their associated FASTQ files (read 1 and read 2)
launch.pbs → a script to submit the job to the HPC cluster
Example index.csv file for nf-core/rnaseq version 3.3:
group,fastq_1,fastq_2,strandedness control_r1,/work/kenna_team/data/raw_data/SRR1039508_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039508_2.fastq.gz,unstranded dex_r1,/work/kenna_team/data/raw_data/SRR1039509_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039509_2.fastq.gz,unstranded control_r2,/work/kenna_team/data/raw_data/SRR1039512_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039512_2.fastq.gz,unstranded dex_r2,/work/kenna_team/data/raw_data/SRR1039513_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039513_2.fastq.gz,unstranded control_r3,/work/kenna_team/data/raw_data/SRR1039516_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039516_2.fastq.gz,unstranded dex_r3,/work/kenna_team/data/raw_data/SRR1039517_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039517_2.fastq.gz,unstranded control_r4,/work/kenna_team/data/raw_data/SRR1039520_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039520_2.fastq.gz,unstranded dex_r4,/work/kenna_team/data/raw_data/SRR1039521_1.fastq.gz,/work/kenna_team/data/raw_data/SRR1039521_2.fastq.gz,unstranded
Example launch.pbs script:
#!/bin/bash -l #PBS -N nfrnaseq #PBS -l select=1:ncpus=2:mem=4gb #PBS -l walltime=24:00:00 #Use the current directory to run the workflow cd $PBS_O_WORKDIR module load java NXF_OPTS='-Xms1g -Xmx4g' #run the nextflow RNA-seq pipeline: nextflow run nf-core/rnaseq -profile singularity -r 3.3 --aligner star_salmon --input index.csv --genome GRCh38 -resume
where:
--aligner Specifies the alignment algorithm to use - available options are 'star_salmon', 'star_rsem', and 'hisat2'. The default option is 'star_salmon'.
more information at:
https://nf-co.re/rnaseq/3.3/usage
More advanced launch.pbs script example:
#!/bin/bash -l #PBS -N nfrnaseq #PBS -l select=1:ncpus=2:mem=4gb #PBS -l walltime=24:00:00 #Use the current directory to run the workflow cd $PBS_O_WORKDIR module load java NXF_OPTS='-Xms1g -Xmx4g' #run the nextflow RNA-seq pipeline: nextflow run nf-core/rnaseq -profile singularity -r 3.3 --aligner star_salmon --input index.csv --genome GRCh38 --clip_r1 10 --clip_r2 10 --three_prime_clip_r1 2 --three_prime_clip_r2 2 --save_trimmed
Session 2 exercises:
Run the nf-core/rnaseq pipeline using the Airway smooth muscle public data (PMID: 24926665. GEO: GSE52778) - aligner option set to ‘star_salmon’
Same as above but aligner option set to ‘star_rsem’
Create a new working folder:
mkdir run1_star_salmon cd run1_star_salmon
Copy index.csv and launch.pbs files to the newly created folder
cp /work/kenna_team/scripts/star_salmon/* .
Check that files were copied into the new working folder
ls -a ./ ../ index.csv launch.pbs #verify the content of index.csv cat index.csv #also check the PBS Pro submission script cat launch.pbs
Run the workflow:
qsub launch.pbs
Monitor the progress of the workflow:
qjobs
or
qstats -u userID
Repeat the above process for ‘star_rsem’
The only variation is copying the index.csv and launch.pbs script. As follows:
cp /work/kenna_team/scripts/star_rsem/* .
Visualizing results
The results generated in the pipeline can be visualized within the ‘results’ folder.
#go to the results folder - note by default all nextflow pipelines show the key outputs within the'results' folder, while the 'work' folders contains all intermediate files generated during execution. cd results #list folders and files ls
example output:
drwxrws--- 2 barrero 4.0K Sep 7 20:05 fastqc/ drwxrws--- 3 barrero 4.0K Sep 7 20:16 trimgalore/ drwxrws--- 3 barrero 23 Sep 9 13:03 multiqc/ drwxrws--- 2 barrero 4.0K Sep 9 13:03 pipeline_info/ drwxrws--- 20 barrero 4.0K Sep 14 23:23 star_rsem/
Access the HPC files from your laptop
Mac laptop (note: need to be connected via VPN)
Open the ‘Finder' window
Click on the search file tab and hit the “Command + K” keys simultaneously
This will open a new window:
Type the above to connect to the shared ‘work' space. To access your personal space replace ‘work’ with 'home’.