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deinterleave FASTQ files
# Usage: < interleaved.fastq f.fastq r.fastq [compress]
# Deinterleaves a FASTQ file of paired reads into two FASTQ
# files specified on the command line. Optionally GZip compresses the output
# FASTQ files using pigz if the 3rd command line argument is the word "compress"
# Can deinterleave 100 million paired reads (200 million total
# reads; a 43Gbyte file), in memory (/dev/shm), in 4m15s (255s)
# Latest code:
# Also see my interleaving script:
# Inspired by Torsten Seemann's blog post:
# Set up some defaults
# If the third argument is the word "compress" then we'll compress the output using pigz
if [[ $3 == "compress" ]]; then
if [[ ${GZIP_OUTPUT} == 0 ]]; then
paste - - - - - - - - | tee >(cut -f 1-4 | tr "\t" "\n" > $1) | cut -f 5-8 | tr "\t" "\n" > $2
paste - - - - - - - - | tee >(cut -f 1-4 | tr "\t" "\n" | pigz --best --processes ${PIGZ_COMPRESSION_THREADS} > $1) | cut -f 5-8 | tr "\t" "\n" | pigz --best --processes ${PIGZ_COMPRESSION_THREADS} > $2
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michaelsilverstein commented Jun 18, 2021

Thanks everyone! Here is how I have been deinterleaving an entire directory of compressed fastq.gz:


# Deinterleave entire directory of compressed .fastq.gz files and re-compress mates

#Usage: indir outdir

mkdir $2

for file in $1/*
        echo $file
        out1=$2/$(basename ${file%.fastq.gz})_R1.fastq.gz
        out2=$2/$(basename ${file%.fastq.gz})_R2.fastq.gz
        pigz --best --processes 16 -dc $file | $out1 $out2 compress

This script will read compressed files from indir, deinterleave them, and save them to outdir with _R1.fastq.gz and _R2.fastq.gz file extensions.

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telatin commented Oct 1, 2021

Hi! SeqFu bundles seqfu interleave and seqfu deinterleave. It's fast (compiled), and provides an easier and less error-prone CLI experience.
If you want to give a try see SeqFu website.

Can be installed via miniconda: conda install -c bioconda seqfu.

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