Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • gwilymh
    Member
    • Dec 2011
    • 72

    #1

    Samtools is failing to phase bam files accurately

    My colleagues and I are assembling transcriptomes from RNASeq data. Our pipeline can (loosely) be outlined as follows:
    1. De novo assembly using all RNASeq read data
    2. Identification of potential coding DNA sequences (CDS)
    3. Alignment of individual RNASeq libraries against CDS sequences to create BAM files
    4. Phasing of BAM files into individual haplotypes
    5. Generate consensus sequence from phased BAM files

    I have been trying to phase the BAM files from a small dataset with only three putative CDS. Looking at the data in Geneious, there are what appear to be very obvious SNPs and heterozygous sites. However, samtools does not sort the polymorphisms into separate BAM files. Rather, each output file seems to contain a random mixture of reads.

    Any suggestions as to why this might be the case?

    NB: BAM files were created in Bowtie2 version 2.2.5
    NB: We are using Samtools version 1.3
    NB: A picture of one of out BAM files is below. There is clearly a T/G and C/A polymorphisms; and the G allele seems to be linked with the A allele. However, Samtools did not separate reads into separate BAM files.

Latest Articles

Collapse

  • SEQadmin2
    Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
    by SEQadmin2



    Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

    Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
    ...
    10-14-2026, 09:55 AM
  • SEQadmin2
    New Genomics Technologies Take Aim at Long-Standing Limits
    by SEQadmin2


    Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

    We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
    ...
    09-28-2026, 10:25 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 10-22-2026, 11:13 AM
0 responses
48 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-22-2026, 10:48 AM
0 responses
36 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-15-2026, 10:52 AM
0 responses
46 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-14-2026, 05:39 AM
0 responses
46 views
0 reactions
Last Post SEQadmin2  
Working...