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  • HTSeq output file

    Hello , I ran a bam file and a sam file through HTSeq for the read counts.
    The following the output of the BAM file with no numeric value for the read counts:
    Code:
    krishna@dntdaretouchit:/mnt/e/cannon$ head countread.text XF:Z:__ambiguous[ENSMUSG00000098178.1+ENSMUSG00000106106.2] XF:Z:__ambiguous[ENSMUSG00000098178.1+ENSMUSG00000106106.2] XF:Z:__alignment_not_unique XF:Z:__alignment_not_unique XF:Z:__alignment_not_unique XF:Z:__alignment_not_unique XF:Z:__no_feature XF:Z:__no_feature XF:Z:__alignment_not_unique XF:Z:__alignment_not_unique
    And this is the output format from the SAM file:
    Code:
    chr1    3206084 255     1S139M  =       3206084 -139    NTACAGTTAACCAACTTATACAGTTAACCAACTCCTACACTAGGTTCCTGAGCATTTCCTTAAACTTGCTAGTTCTGGTTTCCTGGCATGTGAGAGTAAGTCACATGGTAGGAGGCTGCCTTTCTATCJJJFJJFJJJAJFJJJFJFFJFJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJJJFJJJJJA<<JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJJFFJJJJJFJFJJJJJJJJFJJJJJJJJJJJJJJAFAAA     NH:i:1  HI:i:1  AS:i:276        nM:i:0  XF:Z:ENSMUSG00000051951.5
    GWNJ-0965:181:GW180227920:7:2124:9881:65265     163     chr1    3206084 255     139M1S  =       3206084 139     TACAGTTAACCAACTTATACAGTTAACCAACTCCTACACTAGGTTCCTGAGCATTTCCTTAAACTTGCTAGTTCTGGTTTCCTGGCATGTGAGAGTAAGTCACATGGTAGGAGGCTGCCTTTCTATCATTCAATTTTAGN
    I am having hard time to interpret these file for the count numbers. The output from the BAM file looks absurd.
    Last edited by GenoMax; 03-25-2019, 08:16 AM.

  • #2
    I think we need more information.
    What is the design of your study? why HTSeq?
    I never worked with HTSeq before but i have experience with edgeR, DESeq2 and NOISeq, but i can be more useful if i have more information about your design.

    The counts can be extract in R with Rsamtools (look for index function) or in terminal by samtools idsxstats [bam] > [export_to_file]

    Comment


    • #3
      Originally posted by kuldeep View Post
      And this is the output format from the SAM file:
      Code:
      chr1    3206084 255     1S139M  =       3206084 -139    NTACAGTTAACCAACTTATACAGTTAACCAACTCCTACACTAGGTTCCTGAGCATTTCCTTAAACTTGCTAGTTCTGGTTTCCTGGCATGTGAGAGTAAGTCACATGGTAGGAGGCTGCCTTTCTATCJJJFJJFJJJAJFJJJFJFFJFJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJJJFJJJJJA<<JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJJFFJJJJJFJFJJJJJJJJFJJJJJJJJJJJJJJAFAAA     NH:i:1  HI:i:1  AS:i:276        nM:i:0  XF:Z:ENSMUSG00000051951.5
      GWNJ-0965:181:GW180227920:7:2124:9881:65265     163     chr1    3206084 255     139M1S  =       3206084 139     TACAGTTAACCAACTTATACAGTTAACCAACTCCTACACTAGGTTCCTGAGCATTTCCTTAAACTTGCTAGTTCTGGTTTCCTGGCATGTGAGAGTAAGTCACATGGTAGGAGGCTGCCTTTCTATCATTCAATTTTAGN
      It looks like you copy/pasted partial lines in this example. Does your example really start with "chr 1". If so you may have a corrupt SAM file.

      Comment

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