Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • BWA insert size distribution too large

    I'm trying to align pe reads to our reference genome (Tribolium), which has a couple thousand unmapped contigs. The reference has 10 chromosomes , each around 10 - 30 Mb in size, while the rest of the contigs are around 5kb in size.

    I ran this command on the reference with unknown contigs:
    bwa mem -k 5 -t 10 -Ma Ref.fa reads_1.fastq reads_2.fastq > map.sam

    [M::main_mem] read 1164592 sequences (100000019 bp)...
    [M::mem_pestat] # candidate unique pairs for (FF, FR, RF, RR): (23, 23, 12, 19)
    [M::mem_pestat] analyzing insert size distribution for orientation FF...
    [M::mem_pestat] (25, 50, 75) percentile: (2763, 6434, 8256)
    [M::mem_pestat] low and high boundaries for computing mean and std.dev: (1, 19242)
    [M::mem_pestat] mean and std.dev: (5558.52, 3049.04)
    [M::mem_pestat] low and high boundaries for proper pairs: (1, 24735)
    and without the unknown contains:
    bwa mem -k 5 -t 10 -Ma Ref_noUnknown.fa reads_1.fastq reads_2.fastq > map_noUnknown.sam

    [M::main_mem] read 1164592 sequences (100000019 bp)...
    [M::mem_pestat] # candidate unique pairs for (FF, FR, RF, RR): (64, 83, 49, 61)
    [M::mem_pestat] analyzing insert size distribution for orientation FF...
    [M::mem_pestat] (25, 50, 75) percentile: (69, 198, 546)
    [M::mem_pestat] low and high boundaries for computing mean and std.dev: (1, 1500)
    [M::mem_pestat] mean and std.dev: (244.98, 238.31)
    [M::mem_pestat] low and high boundaries for proper pairs: (1, 1977)
    Why is the insert size distribution different between these alignments? How can the insert size distribution be so large for the alignment that contains 2000+ contigs?

  • #2
    Problem solved.

    I ran fastq_quality_trimmer on my raw reads, and the output PE files did not match up, causing large inferred insert sizes.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Essential Discoveries and Tools in Epitranscriptomics
      by seqadmin


      The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist on Modified Bases...
      Yesterday, 07:01 AM
    • 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

    ad_right_rmr

    Collapse

    News

    Collapse

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