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  • MiSeq fastQ quality scores

    I have fastQ files as output from MiSeq, 300bp directional reads. I would like to plot the distribution of quality scores for all bases (using Perl, Gnuplot or R).

    I can't find definite information on how to interpret the quality string in fastQ-files coming from MiSeq.

    Could anyone point me to this info? Thanks!

  • #2
    You might like to try FastQC : http://www.bioinformatics.babraham.a...ojects/fastqc/

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    • #3
      Originally posted by Azazel View Post
      I have fastQ files as output from MiSeq, 300bp directional reads. I would like to plot the distribution of quality scores for all bases (using Perl, Gnuplot or R).

      I can't find definite information on how to interpret the quality string in fastQ-files coming from MiSeq.

      Could anyone point me to this info? Thanks!
      Information about the FASTQ format can be found here: http://en.wikipedia.org/wiki/FASTQ_format With MiSeq you are looking at quality scores in the "L" (Illumina 1.8+) format as referred to in the Wikipedia article.

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      • #4
        Originally posted by GenoMax View Post
        Information about the FASTQ format can be found here: http://en.wikipedia.org/wiki/FASTQ_format With MiSeq you are looking at quality scores in the "L" (Illumina 1.8+) format as referred to in the Wikipedia article.
        @GenoMax: Thanks. I have read the Wikipedia article previously, but I find it ambiguously formulated and I don't know what the "Illumina pipeline" is. As a reader of that article, I know I using MiSeq now but it looks like this was writen only with respect to HiSeq.

        I am not trying to argue, just pointing out that the Wikipedia article is a bit unclear on that point.

        @kadircanir: Thanks for pointing out this program! It is excellent.

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        • #5
          PRINCESEQ can produce decent graphs

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          • #6
            You can also use the FASTX toolkit (just remember to use the -Q33 setting!) to produce quality graphs over cycle. With 300 cycles the cycle numbers will all moosh together a bit though.

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            • #7
              Originally posted by kadircaner View Post
              FastQC works well

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              • #8
                Hopefully, the plots generated by BaseSpace for the MiSeq output will be exportable into plots soon, that might ease some of this extra effort a bit!

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