Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • HTseq - count - high ambiguous count rate and reads with missing mate encountered

    Hi. I'm using HTseq to produce some count read files that I can then use for NOIseq and edgeR. I used trimmomatic to trim my reads in paired end mode and merged the paired ends in tophat2 using

    Code:
    /home/user/Downloads/tophat2/tophat -p 4 -o /home/user/Desktop/sam/trimmomatic_out/2B82 --mate-inner-dist 100 --library-type fr-firststrand /home/user/Downloads/bowtie2/example/index/DinoAnt /home/user/Desktop/sam/trimmomatic_out/2B82/output_forward_paired.fastq.gz /home/user/Desktop/sam/trimmomatic_out/2B82/output_reverse_paired.fastq.gz
    I then sorted them using the samtools sort function. When I run the resulting bam files through HTseq count function, I get a really high percentage (95%) of reads with missing mate pair and also around 65% amiguous feature rate.

    I used this command and ran it on a gtf file I produced myself in cuffmerge from the RNAseq data I collected.

    Code:
    htseq-count -m union -s no -t exon -f bam -i gene_id /home/user/Desktop/sam/trimmomatic_out/10G87/accepted_hits.sorted.bam /home/user/Desktop/new_dino_merged.gtf
    Can anyone help me why I am getting such high ambiguous count rate and such a high percentage of missing mate pairs?

    Thanks
    Last edited by 4galaxy7; 12-14-2015, 05:19 AM.

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...
    Today, 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
37 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 10:19 PM
0 responses
41 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 09:21 AM
0 responses
35 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-04-2024, 09:00 AM
0 responses
54 views
0 likes
Last Post seqadmin  
Working...
X