Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • bwa outputs bam with less reads than in input fastq

    I downloaded a publicly available bam file and ran samtools flagstat on it. The output is here:

    482805326 + 0 in total (QC-passed reads + QC-failed reads)
    0 + 0 duplicates
    235328156 + 0 mapped (48.74%:nan%)
    482805326 + 0 paired in sequencing
    241402663 + 0 read1
    241402663 + 0 read2
    235328156 + 0 properly paired (48.74%:nan%)
    235328156 + 0 with itself and mate mapped
    0 + 0 singletons (0.00%:nan%)
    0 + 0 with mate mapped to a different chr
    0 + 0 with mate mapped to a different chr (mapQ>=5)

    Next, I extracted reads from the bam into two fasta files. The sum of reads in both was 482805326, so equal as in the bam. I aligned the fastq files again (bwa aln and bwa sampe), and ran samtools flagstat on the resulting bam. Here is the output:

    387280125 + 0 in total (QC-passed reads + QC-failed reads)
    0 + 0 duplicates
    387280125 + 0 mapped (100.00%:nan%)
    387280125 + 0 paired in sequencing
    194031131 + 0 read1
    193248994 + 0 read2
    375578347 + 0 properly paired (96.98%:nan%)
    381759377 + 0 with itself and mate mapped
    5520748 + 0 singletons (1.43%:nan%)
    5108005 + 0 with mate mapped to a different chr
    4124551 + 0 with mate mapped to a different chr (mapQ>=5)

    And what I don't understand is where is almost 100M reads gone? I haven't found any information on bwa removing unmapped/bad quality/duplicate reads, so I am wondering. Any hints?

    I am using bwa 0.5.9-r16
    Last edited by seru; 03-06-2012, 03:07 AM. Reason: added tool version

  • #2
    Same here

    I have this same problem. Did you ever figure out how to fix it?

    Update: The missing reads in the bam file were due to the following error:
    [bwa_sai2sam_pe_core] print alignments... [E::sam_parse1] CIGAR and query sequence are of different length
    [W::sam_read1] parse error at line 11261615
    [main_samview] truncated file.

    All of this happened after I switched to a new reference genome (mm10). I'm not getting the error with mm9. I don't have a good explanation but I think that the version of mm10 I tried to use must have been somehow corrupted.
    Last edited by Guest; 11-05-2016, 03:37 AM.

    Comment


    • #3
      Note the "100%" mapped metric, which means that the unmapped reads were excluded from the output.

      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...
        04-22-2024, 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
      59 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      57 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      51 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      56 views
      0 likes
      Last Post seqadmin  
      Working...
      X