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  • crh
    Member
    • Dec 2009
    • 46

    illumina to fastq

    I'm new to working with NGS and we have some illumina reads we'd like to convert to fastq.
    I've read the posts and tried to run the script qseq2fastq.pl on the samples w/o success.

    the files have the following format which doesn't look quite like qseq:

    USI-EAS39:1:1:2:1362#0/1:CTGGNTTCCACAGGCACATAGCCAAACCGGTGCCT:32 32 29 24 4 28 33 32 32 34 32 34 32 30 33 33 32 33 33 33 33 32 33 29 30 33 33 33 28 21 26 30 30 33 30

    Appreciate any conversions ideas.

    Charles
  • simonandrews
    Simon Andrews
    • May 2009
    • 870

    #2
    The script below should convert this format of data into a fastq file with Illlumina 1.5 quality encoding (you can change the offset value on line 20 if you'd prefer Sanger format).

    Code:
    #!/usr/bin/perl
    use warnings;
    use strict;
    
    my ($infile,$outfile) = @ARGV;
    
    die "Must specify input and output filename\n" unless ($outfile);
    
    open (IN,$infile) or die $!;
    open (OUT,'>',$outfile) or die $!;
    
    while (<IN>) {
      chomp;
      my @sections = split(/:/);
    
      my @qualities = split(/\s+/,$sections[-1]);
    
      my $quality_string = '';
    
      $quality_string .= chr($_ + 64) foreach (@qualities);
    
      my $seq_id = join(":",@sections[0..4]);
      print OUT '@',$seq_id,"\n",$sections[5],"\n+",$seq_id,"\n",$quality_string,"\n";
    
    }
    
    close OUT or die $!;

    Comment

    • crh
      Member
      • Dec 2009
      • 46

      #3
      Thanks Simon,

      what should the offset be for sanger fastq?

      Charles

      Comment

      • simonandrews
        Simon Andrews
        • May 2009
        • 870

        #4
        Originally posted by crh View Post
        what should the offset be for sanger fastq?
        33 (apparently I need to add some more text otherwise the forum won't allow the post...)

        Comment

        • crh
          Member
          • Dec 2009
          • 46

          #5
          Thanks again Simon - works fine.

          Is the original format I have 'Gerald'?

          Charles

          Comment

          • ETHANol
            Senior Member
            • Feb 2010
            • 308

            #6
            Biopython Seq I/O does these conversions. Pretty easy to use and with pretty good documentation.
            --------------
            Ethan

            Comment

            • kmcarr
              Senior Member
              • May 2008
              • 1181

              #7
              Originally posted by crh View Post
              Thanks again Simon - works fine.

              Is the original format I have 'Gerald'?

              Charles
              It's a very old Solexa format called SCARF (Solexa Compact ASCII Read Format). It's not used anymore. Is this data really old?
              Last edited by kmcarr; 04-08-2011, 12:14 PM. Reason: Expand acronym

              Comment

              • crh
                Member
                • Dec 2009
                • 46

                #8
                I've used bioperl mostly, I'll check if they have seq conversions for illumina formats.
                regarding the reads, I believe they are ~2 yrs old now.

                thanks

                Comment

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