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  • Sorted bamfiles have increased size!!

    BACKGROUND:
    I have RNA-Seq data from Illumina platform.
    I aligned the reads post-QC onto hg19 bowtie index using tophat.
    Now I wished to get the raw count of reads mapped to each gene using Ht-seq-count.

    WHAT I DID:
    However the HT-Seq reports error that it is unable to find the mate pair and asks whether the sam file is properly sorted.
    Hence, after reading posts on the problem. I sorted my bam files using samtoools.
    samtools sort -n accepted_hits.bam accepted_hits_bam_sorted

    CONCERN:
    Now the sorted BAM file has increased in size and I wish to know why his has happened.
    Though it might not be but seems highly unintuitive.

  • #2
    That's not surprising. Remember that BAM files are compressed, with the compression algorithm being more efficient the more similar neighbouring reads are (i.e., coordinate-sorted files should compress to a greater extent).

    Comment


    • #3
      Thanks dpryan.
      Just wanted to confirm if something went horribly wrong.
      I'll now be following your earlier reply on a different post to run HTSeq

      Comment


      • #4
        You sorted by read name (-n), but the default sorting by mapping position puts similar reads together, and therefore their sequence data compresses well, so coordinate sorted BAM files are usually smaller.

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