Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • BWA mem reporting poor alignments

    I've been using bwa mem to align illumina reads to contaminant genomic databases (ribosomal RNAs, gbbct, etc). I've noticed unusually high percentages of reads mapping to my contaminants (10% or more when I expected relatively little contamination) and suspiciously high percentages of reads mapping to multiple contaminant databases (human as well as gbbct, for instance). I blast-ed a few of these "contaminant" bacterial reads online, and they were showing relatively poor alignments to bacterial sequences (e.g., 30-40% of the read mapping with a few mismatches).

    Right now I'm using the default parameters, which obviously isn't working. I'm wondering if someone could give me some advice on how to tweak things to restrict myself to better alignments.

  • #2
    You could filter on mapping quality, e.g. by piping through samtools a la

    bwa mem ref fastq1 fastq2 | samtools view -S -q30 -

    Comment


    • #3
      That sounds like a good idea. Do you know the range in the quality scores? What's a reasonable cutoff to ask for? Is -q 30 what you would suggest?

      Comment


      • #4
        And one more thing ...
        I've read that BWA gives ambiguously mapped reads a quality of 0. Would I have to pull those reads out separately somehow?

        Comment

        Latest Articles

        Collapse

        • seqadmin
          Current Approaches to Protein Sequencing
          by seqadmin


          Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
          04-04-2024, 04:25 PM
        • seqadmin
          Strategies for Sequencing Challenging Samples
          by seqadmin


          Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
          03-22-2024, 06:39 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by seqadmin, 04-11-2024, 12:08 PM
        0 responses
        18 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-10-2024, 10:19 PM
        0 responses
        22 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-10-2024, 09:21 AM
        0 responses
        16 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-04-2024, 09:00 AM
        0 responses
        47 views
        0 likes
        Last Post seqadmin  
        Working...
        X