Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • NGSfan
    Senior Member
    • Apr 2009
    • 181

    #1

    Too many mismatches aligned by BFAST?

    Just found an interesting result from one of my BFAST alignments.

    I have two replicates of an target enrichment (exome) sequencing run.

    In one of the replicates I have pretty clean reads (MQ = 255) and alignments

    In the other replicate, I see 12 reads that look out of place (with MQ values ~150). The quality scores per bp are high (~30-38) so it's not sequencing error perhaps, yet these reads don't look like they belong there given the extremely high number of mismatches.

    1) How do I prevent/filter out these sorts of alignments? I'm using -a 3 in the BFAST postprocess (uniquely select the best scoring alignment).

    2) Also, any one have any theories as to how these reads came about?? What kind of artifact is this? alignment or PCR?

    Thanks for any ideas/thoughts/suggestions!

    Since there is a 97kb limit for images, I've got my IGV screenshot here:
    SendSpace lets you transfer large files quickly and securely for free. Enjoy hassle-free file sharing today.
    Last edited by NGSfan; 11-25-2010, 04:12 AM. Reason: cannot upload image
  • NGSfan
    Senior Member
    • Apr 2009
    • 181

    #2
    the other interesting thing is that the pair mates are in the opposite direction.

    Normal pair mates appear as -----> <------
    While these highly mismatched reads, the paired mates are in the opposite orientation <------- ------->

    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
    • SEQadmin2
      Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
      by SEQadmin2



      Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
      ...
      07-09-2026, 11:10 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, Yesterday, 10:13 AM
    0 responses
    13 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-31-2026, 02:55 AM
    0 responses
    25 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-24-2026, 12:17 PM
    0 responses
    19 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-23-2026, 11:41 AM
    0 responses
    18 views
    0 reactions
    Last Post SEQadmin2  
    Working...