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FastQC with multiple FASTQ files

I have received 384 fastq.gz files. These come from paired-end sequencing so I have 2 files per patient so 192 patients. I am new to NGS data analysis and I wish to start using FastQC. What would be the best way to proceed?

  • I know FastQC can be run graphically but presumably, with that many samples, it would be best to use the command line..
  • I read some places that merging all samples into a single (or 2 with paired-end) files might be the solution. Is that recommended? Or should I just use simple bash scripting in like below (or something similar)?

    for i in *fastqc.gz do bsub < fastqc_script_with_commands.sh done

I guess I'm just curious if there is a convention of merging fastq files or keeping them separate (1 or 2 per sample).

Thanks

ngs fastqc multiple

2 answers

fastqc.sh:

#!/usr/bin/env bash
RUN_PATH=$1
cd $RUN_PATH
for file in $(ls $RUN_PATH)
do
    SAMPLE=`basename $file`
    fastqc -t 5 ${SAMPLE} -o /path/to/where/you/want/outputs
done
$./fastqc.sh /path/to/fastqs/

If you are running this on a cluster, just add qsub or bsub before the fastqs line (we have ibm):

That line would become:

bsub -P project -q queuename -n 1 -R "rusage[mem=2000]" fastqc -t 5 ${SAMPLE} -o /path/to/where/you/want/outputs

Change the queuename to the actual name of the queue you submit jobs to.

You would want to do the QC for files individually. When run on the command line with -o option FastQC will write the result files to that directory. A bash loop would work. You can look into MultiQC to aggregate all results.

+1 for MultiQC, I dont even bother to look at the individual output metrics anymore

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