Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Can or should we perform feature counting with `bedtools intersect -c`?

    I have the following BAM file and a annotation file.
    What I want to do is to calculate reads count in
    I can do it with the following three methods.


    Below is the code

    Code:
        #!/bin/bash
        INPUT_BAM=my_paired_end_alignment.bam
        ANNNOT_BED=peak_annot.bed
        ANNNOT_GTF=peak_annot.gtf
        
        # Temporary file
        ALIGNMENT_BED=my_paired_end_alignment.bed
        
        #-----------
        # Method 1
        #-----------
        FINAL_COUNT_METHOD1=output_method1.txt
        bedtools bamtobed -bedpe -i $INPUT_BAM > $ALIGNMENT_BED
        bedtools intersect -a $ANNOT_BED -b $ALIGNMENT_BED -c > $FINAL_COUNT_METHOD1
        
        #-----------
        # Method 2
        #-----------
        FINAL_COUNT_METHOD2=output_method2.txt
        bedtools multicov -bams $INPUT_BAM -bed $ANNOT_BED > $FINAL_COUNT_METHOD2
        
        #-----------
        # Method 3
        #-----------
        featureCounts  -a $ANNOT_GTF -o counts.txt $INPUT_BAM
    The result given by Method1 is so different from Method2 (totally uncorrelated),
    whereas Method2 is highly correlated with Method3.

    My question is Method1 the right way to count feature? If not why?
    Should we use Method2 or Method3 instead?

  • #2
    For reference cross-posted: https://www.biostars.org/p/270366/

    Comment

    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...
      05-06-2024, 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, 06:57 AM
    0 responses
    12 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 05-06-2024, 07:17 AM
    0 responses
    16 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
    24 views
    0 likes
    Last Post seqadmin  
    Working...
    X