Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Removing Duplicates Scenario Exome Resequencing

    I am rethinking my well-established pipeline and trying to tweek it a bit and came across some controversy on the use of remove duplicates. I would like to propose an example scenario and my current understanding of Picard Tool's Mark Dups:

    At a given 5' loci (chr12:144230), two alleles are present (assume these are the first three nucleotides of 90bp reads):
    1) GAC #reads = 30 Highest Q score= 25
    2) GAA #reads = 8 Highest Q score= 26

    It is possible that some of the 30 reads of allele 1 are due to strand bias or pcr dup. and if visualizing this on an aligner, I would have some suspicion that the locus is heterozygous.

    If Mark Duplicates: Allele 2 is kept due to highest Q score, and thus there is loss of heterozygosity

    If no duplicate removal: Both alleles are kept, but the ration is not close enough to 50% to be called a het.

    Is my very rudimentary understanding of this tool correct and, if so, are there any other solutions?
    Does anyone have recommendations based on the fact that this particular project is exome resequencing at 50-75x average depth?)

  • #2
    With 75x coverage and I'm assuming paired end reads that has a size distribution with a standard deviation of at least 30bp, it is very unlikely for you to get multiple reads to pile up at the same exact locations without them being PCR duplicates. I would remove them for SNP calling.

    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
    9 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, Yesterday, 01:02 PM
    0 responses
    8 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 09-18-2024, 06:39 AM
    0 responses
    10 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 09-11-2024, 02:44 PM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Working...
    X