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Sequence Length Distribution From A Fastq File

Hi all,

Any script available for doing sequence length disrtibution of fastq file?

Thanks, Deepthi

sequence length fastq

I'm surprised anyone answered this, given that you present no evidence of having looked for a solution yourself.

Good point: I'm as a consequence surprised by myself...

Maybe just let the community decide on whether a question is worthy of answering or not (unless of course it is completely unsuitable for Biostar). To most of us who have been at this a while his question is trivial, but to him... Lets not make people feel bad about asking questions.

I don't care if questions seem "trivial". I do care about questions that begin "is there any script" - it simply implies that they have not even tried a Web search before asking.

I got here 7 years later from having the same question and googling...honestly I come across so many posts like this when I google and there's always comments shaming OP. I for one really appreciate the people that ask questions that others judge.

Hello,

I am trying to do something similar, were I want to determine both the read length and how many reads are in my fastq file. Here is my code:

gunzip -c SRR1060507_1.fastq.gz|awk 'NR%4==2{printlength($0)}'|uniq -c

But I keep getting the following error:

awk: cmd. line:1: (FILENAME=- FNR=2) fatal: function `printlength' not defined

I'm not sure what I've done incorrectly? I also tried Frederic's code above and although I got that to run, its not exactly the output I'm seeking, I should be returning something like 2420797 100

Any help would be super appreciated!

Between print and length should be at least one space. If you want to compute the number of reads simultaneously, use something like Frèdèric's approach

awk 'NR%4 == 2 {lengths[length($0)]++ ; counter++} END {for (l in lengths) {print l, lengths[l]}; print "total reads: " counter}' file.fastq

Thanks! This was a winner.

Anyone know how to divide reads to create two plots, one for redundant and one for unique, like this: example plot

7 answers

Here is a solution using awk:

awk 'NR%4 == 2 {lengths[length($0)]++} END {for (l in lengths) {print l, lengths[l]}}' file.fastq

It reads like this: every second line in every group of 4 lines (the sequence line), measure the length of the sequence and increment the array cell corresponding to that length. When all lines have been read, loop over the array to print its content. In awk, arrays and array cells are initialized when they are called for the first time, no need to initialize them before.

Thanks for teaching me the NR%4 line grouping method: pretty useful indeed when dealing with fastq files.

Another awk solution, very similar to Frédéric's:

cat reads.fastq | awk '{if(NR%4==2) print length($1)}' | sort -n | uniq -c > read_length.txt

And to quickly obtain a graph in R:

reads<-read.csv(file="read_length.txt", sep="", header=FALSE)
plot (reads$V2,reads$V1,type="l",xlab="read length",ylab="occurences",col="blue")

no need to use cat

With the cat approach you can also zcat reads.fastq.gz | ...

You can use FASTQC. It gives you all the details you need, including the quality, overrepresented sequences, enriched k-mers etc.

You could do that but you may want to use NanoPlot instead. It will generate stats that are specific for nanopore sequencing and thus more useful.

Another possibility, from BBMap:

readlength.sh in=reads.fq out=histogram.txt

The default is 10bp bins with a max of 80kbp, but those can be configured (run the shellscript with no arguments for details). It's very fast, and handles fastq/fasta/sam/bam; raw/gzip/bzip2.

Really useful! Only if I want to parse the output (e.g. pipe it into grep) I have to first suppress the stderr message: Processed XY reads. Time: Z seconds. Might it be more convenient to have this info also in stdout?

Save it as fastq_lenght.py and run it :)

#!/usr/bin/env python
#-*- coding: UTF-8 -*-
import sys
######################################################################################
syntax = '''
------------------------------------------------------------------------------------
Usage: python fastq_lenght.py file.fastq 
result: .txt file same name as input name plus "_lenghts.txt" 
------------------------------------------------------------------------------------
'''
######################################################################################

if len(sys.argv) != 2:
    print syntax
    sys.exit()

######################################################################################

fastq_file = open(sys.argv[1], 'r')
prefix = sys.argv[1].split('.')[0]
outfile = open(prefix + '_' + 'lenghts.txt', 'w')
seq = ''
for line in fastq_file:
    line = line.rstrip('\n')
    if line.startswith('@'):
        if seq:
            outfile.write(name + '\t' + str(len(seq)) + '\n')
            seq = ""
        name = line 
    else:
        seq = line 
outfile.write(name + '\t' + str(len(seq)) + '\n')
fastq_file.close()
outfile.close()

print '\n' + '\t' + 'File: ' + prefix + '_' + 'lenghts.txt has been created...'

You can do this using fastx toolkit, especially, you might be interested in this program:

fastx_nucleotide_distribution_graph.sh

and maybe:

fastx_quality_stats
fastq_quality_boxplot_graph.sh

i think this does not provide length distribution information

Use this post as a help Fastq Quality Read and Score Length Check and apply a few basic unix commands like sort and uniq to get the frequency.

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