Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • Extract reads from paired-end fastq based on specific adapters with bbduk

    Hello everyone, I am using bbduk.sh (from bbmap toolkit) to extract reads from paired-end fastq files based on the presence of specific adapters in the 5' of the sequence in the "_1" fastq file.

    I am using this command:

    Code:
    ./bbmap/bbduk.sh -Xmx1g in1=reads_1.fastq.gz in2=reads_2.fastq.gz outm1=matched1.fastq.gz outm2=matched2.fastq.gz literal=AAACCTGAGAAACCTA k=16 hdist=0 -rcomp=f
    The problem is that other that the correct reads, the output file contains also other reads which do not include the adapter sequence, es:

    # from reads_1.fastq.gz
    @SRR9262917.232075 232075/1
    GCATGCGAGTAGCGGTGGTTCTTATA
    +
    FFFFFFFFFFFFFFFFFFFFFFFFFF

    # from reads_2.fastq.gz
    @SRR9262917.232075 232075/2
    AAGCAGTGGTATCAACGCAGAGTACATGGGATTCCATAGCCCTGTGGTTTTTATAGATCTTGTAAACCCCAAACCTGGGAAACCTAGTGGC
    +
    FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFF,FFFFFFFFF

    Does anyone know why this may be happening and how to avoid this?

    Thanks in advance.

  • #2
    You could add a "restrictleft=N" N=certain number of bases to look only in that area. Also adding "minlength=N" will exclude small reads like the first example. Also try setting k to something smaller (8) so it has better chances of matching correctly.

    I hope "-rcomp=f" is a typo. There should be no - at beginning.
    Last edited by GenoMax; 11-08-2019, 06:44 AM.

    Comment


    • #3
      Thank you! That worked

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Understanding Genetic Influence on Infectious Disease
        by seqadmin




        During the COVID-19 pandemic, scientists observed that while some individuals experienced severe illness when infected with SARS-CoV-2, others were barely affected. These disparities left researchers and clinicians wondering what causes the wide variations in response to viral infections and what role genetics plays.

        Jean-Laurent Casanova, M.D., Ph.D., Professor at Rockefeller University, is a leading expert in this crossover between genetics and infectious...
        09-09-2024, 10:59 AM
      • seqadmin
        Addressing Off-Target Effects in CRISPR Technologies
        by seqadmin






        The first FDA-approved CRISPR-based therapy marked the transition of therapeutic gene editing from a dream to reality1. CRISPR technologies have streamlined gene editing, and CRISPR screens have become an important approach for identifying genes involved in disease processes2. This technique introduces targeted mutations across numerous genes, enabling large-scale identification of gene functions, interactions, and pathways3. Identifying the full range...
        08-27-2024, 04:44 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, Today, 06:25 AM
      0 responses
      13 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, Yesterday, 01:02 PM
      0 responses
      12 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 09-18-2024, 06:39 AM
      0 responses
      14 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 09-11-2024, 02:44 PM
      0 responses
      14 views
      0 likes
      Last Post seqadmin  
      Working...
      X