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Prior running the nf-core/sarek pipeline with real data, we will first run a test with sample data to make sure the pipeline runs properly.

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Code Block
mkdir -p $HOME/workshop/sarek/scripts
cp /work/training/sarek/scripts/* $HOME/workshop/sarek/scripts/ ls -l $HOME/workshop/sarek/scripts/
  • Line 1: The -p indicates create 'parental directories as required. Thus the line 1 command creates both /workshop/ and the subfolder /workshop/sarek/scripts/

  • Line 2: Copies all files from /work/datasets/workshop/scripts/ as noted by an asterisk to the newly created folder $HOME/workshop/sarek/scripts/

Create folders for running the nf-core/sarek pipeline

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Code Block
mkdir $HOME/workshop/sarek/run1_test
mkdir $HOME/workshop/sarek/run2_trio
mkdir $HOME/workshop/sarek/run3_liver
cd $HOME/workshop/sarek
  • Lines 1-3: create sub-folders for each exercise

  • Line 4: change the directory to the folder “run1_test”

Exercise 1: Running a test with nf-core sample data

First, let’s assess the execution of the nf-core/rnaseq pipeline by running a test using sample data.

Copy the launch_nf-core_RNAseqsarek_test.pbs to the working directory

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#!/bin/bash -l

#PBS -N nfsarek_run1_test

#PBS -l walltime=48:00:00

#PBS -l select=1:ncpus=1:mem=5gb

cd $PBS_O_WORKDIR

NXF_OPTS='-Xms1g -Xmx4g'

module load java

#specify the nextflow version to use to run the workflow

export NXF_VER=23.10.1

#run the sarek pipeline

nextflow run nf-core/sarek \

        -r 3.3.2 \

        -profile test,singularity \

        --outdir ./results

  • nextflow command: nextflow run

  • pipeline name: nf-core/sarek

  • pipeline version: -r 3.3.2

  • container type and sample data: -profile test,singularity

  • output directory: --outdir results

Submitting the job

Submit the test job to the HPC cluster as follows:

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Code Block
results/
├── csv
│   ├── markduplicates.csv
│   ├── markduplicates_no_table.csv
│   ├── recalibrated.csv
│   └── variantcalled.csv
├── multiqc
│   ├── multiqc_data
│   ├── multiqc_plots
│   └── multiqc_report.html
├── pipeline_info
│   ├── execution_report_2024-05-08_15-28-38.html
│   ├── execution_timeline_2024-05-08_15-28-38.html
│   ├── execution_trace_2024-05-08_15-28-38.txt
│   ├── params_2024-05-08_15-41-30.json
│   ├── pipeline_dag_2024-05-08_15-28-38.html
│   └── software_versions.yml
├── preprocessing
│   ├── markduplicates
│   ├── recalibrated
│   └── recal_table
├── reports
│   ├── bcftools
│   ├── fastqc
│   ├── markduplicates
│   ├── mosdepth
│   ├── samtools
│   └── vcftools
├── tabix
│   ├── genome.bed.gz
│   └── genome.bed.gz.tbi
└── variant_calling
    └── strelka

Once the pipeline has finished running - Assess the QC report:

NOTE: To proceed, you need to be on QUT’s WiFi network or signed via VPN.

To browse the working folder in the HPC type in the file finder:

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