Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • crh
    Member
    • Dec 2009
    • 46

    #1

    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

                Latest Articles

                Collapse

                • SEQadmin2
                  16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
                  by SEQadmin2




                  NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification, detection...
                  Yesterday, 06:46 AM
                • SEQadmin2
                  Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
                  by SEQadmin2



                  Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

                  Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
                  ...
                  10-14-2026, 09:55 AM

                ad_right_rmr

                Collapse

                News

                Collapse

                Topics Statistics Last Post
                Started by SEQadmin2, 10-28-2026, 09:57 AM
                0 responses
                26 views
                0 reactions
                Last Post SEQadmin2  
                Started by SEQadmin2, 10-28-2026, 09:51 AM
                0 responses
                17 views
                0 reactions
                Last Post SEQadmin2  
                Started by SEQadmin2, 10-22-2026, 11:13 AM
                0 responses
                71 views
                0 reactions
                Last Post SEQadmin2  
                Started by SEQadmin2, 10-22-2026, 10:48 AM
                0 responses
                48 views
                0 reactions
                Last Post SEQadmin2  
                Working...