Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • CarnifexRex
    Junior Member
    • Sep 2018
    • 4

    #1

    Adaper sequence to Read fastq

    Hello,

    Fairly new to bioinformatics and first time posting, hopefully I sound halfway intelligent

    My FASTQs contain adapter sequences in the read name. This adapter sequence includes a UMI. However my postprocessing software requires the UMI to be part of the read. Is there some way of inserting the adapter sequence from the read name to the beginning of the read? I'm guessing I'll also need to insert some bogus quality scores as well.

    To visualize, I need to turn this:
    @M01179:478:000000000-C33YY:1:1101:17276:1520 2:N:0:CTTGTATGTATG
    TTGTCGTTCCTTTCTTTTTGTCTCTTTCCTGTACCTCTAG
    +
    11111333@B1B11AA3A3B1A3B3BFEE333130110A0

    into this:

    @M01179:478:000000000-C33YY:1:1101:17276:1520 2:N:0:CTTGTATGTATG
    CTTGTATGTATGTTGTCGTTCCTTTCTTTTTGTCTCTTTCCTGTACCTCTAG
    +
    AAAAAAAAAAAA11111333@B1B11AA3A3B1A3B3BFEE333130110A0

    This would need to be a scriptable solution, and work on all reads from a High Out-put NextSeq run

    Thanks in advance
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    Are you sure about that? Generally in case of Illumina sequencing index sequences (which are not part of actual reads) are transferred to the fastq header as a part of demultiplexing process. That is what you are probably seeing e.g. "CTTGTATGTATG".

    Comment

    • CarnifexRex
      Junior Member
      • Sep 2018
      • 4

      #3
      That is correct, sequence CTTGTATGTATG is my index sequence. The first 6 bp (CTTGTA) is my sample barcode, the second 6 bp (TGTATG) is a molecular barcode. This second sequence is random, so changes across every read.

      I need to add the entire 12 bp sequence to the beginning of my read in order to utilize the UMI portion of the adapter in my alignment and variant calling software.

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #4
        This may be easier to do by obtaining the index read as a separate fastq file. You will also get real Q-scores for bases in index read when you do that. Stand-alone bcl2fastq allows one to get data in this format. I assume you may be able to do this using BaseSpace as well, if that is what you are using.

        You can then use a program called "Seqkit" (and specifically option "seqkit concat" to concatenate your index read in front of the actual read).

        Comment

        • CarnifexRex
          Junior Member
          • Sep 2018
          • 4

          #5
          Thats fantastic. This confirms by believe that there's a tool for everything.

          This does exactly what I need, with one exception. It calls the entire FastQ into memory, which is a problem when the FastQ files are 20GB each.

          I thought faidx indexing might help, but 1) bcl2fastq outputs gzip not bgzip files, and 2) my input files are FastQ and not FastA. I also thought of trying to stream the job by just providing a list of read names to concatenate using the seqkit common or grep command, but I couldn't get that to work.

          Now I'm thinking of some combination of sort, split and bash loop but I'm still a novice at linux scripting.

          Does anyone have some suggestions on how to process large files using seqkit?

          Thanks

          Comment

          Latest Articles

          Collapse

          • SEQadmin2
            Beyond CRISPR/Cas9: Understand, Choose, and Use the Right Genome Editing Tool
            by SEQadmin2



            CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).

            Despite this, “CRISPR helped turn genome editing from a specialized technique into
            ...
            07-31-2026, 11:01 AM
          • SEQadmin2
            Proteomic Platforms: How to Choose the Right Analytical Strategy to Improve Detection and Clinical Applications
            by SEQadmin2


            Proteomics platforms are evolving rapidly, with advances in mass spectrometry and affinity-based approaches expanding what researchers can detect and at what scale. As the field moves toward deeper proteome coverage and clinical applications, scientists face an increasingly complex landscape of tools. This article will explore how researchers are navigating these choices to find the right platform for their work.

            The systematic characterization of the human proteome has
            ...
            07-20-2026, 11:48 AM

          ad_right_rmr

          Collapse

          News

          Collapse

          Topics Statistics Last Post
          Started by SEQadmin2, 08-06-2026, 07:41 AM
          0 responses
          23 views
          0 reactions
          Last Post SEQadmin2  
          Started by SEQadmin2, 08-03-2026, 10:13 AM
          0 responses
          39 views
          0 reactions
          Last Post SEQadmin2  
          Started by SEQadmin2, 07-31-2026, 02:55 AM
          0 responses
          43 views
          0 reactions
          Last Post SEQadmin2  
          Started by SEQadmin2, 07-24-2026, 12:17 PM
          0 responses
          27 views
          0 reactions
          Last Post SEQadmin2  
          Working...