Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • 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

  • seqadmin
    Recent Advances in Sequencing Analysis Tools
    by seqadmin


    The sequencing world is rapidly changing due to declining costs, enhanced accuracies, and the advent of newer, cutting-edge instruments. Equally important to these developments are improvements in sequencing analysis, a process that converts vast amounts of raw data into a comprehensible and meaningful form. This complex task requires expertise and the right analysis tools. In this article, we highlight the progress and innovation in sequencing analysis by reviewing several of the...
    Yesterday, 07:48 AM
  • 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

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by seqadmin, Yesterday, 07:17 AM
0 responses
11 views
0 likes
Last Post seqadmin  
Started by seqadmin, 05-02-2024, 08:06 AM
0 responses
19 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-30-2024, 12:17 PM
0 responses
20 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-29-2024, 10:49 AM
0 responses
28 views
0 likes
Last Post seqadmin  
Working...
X