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  • htseq-count failure parsing GFF attribute line

    Hi,
    I am trying to generate a read counts file for my ChIP-seq data using htseq-count. Here is what I have done so far:

    1. Call ChIP-seq peaks and output BED file in MACS2
    2. Convert BED to GFF in Galaxy
    2. Run htseq-count with the following command and error:

    Code:
    -bash-4.1$ htseq-count -f bam -s no -t feature M1-cbx5_sorted.bam Rep1_cbx5_merge.gff
    Error occured when processing GFF file (line 1 of file Rep1_cbx5_merge.gff):
      Failure parsing GFF attribute line
      [Exception type: ValueError, raised in __init__.py:164]
    I suspect that there is something wrong with my GFF input, but I am not sure what.

    Here is what my GFF file looks like:
    Code:
    -bash-4.1$ head Rep1_cbx5_merge_edit.gff
    chr1	bed2gff	feature	823401	825999	0	+	.	group1;
    chr1	bed2gff	feature	927001	928999	0	+	.	group2;
    chr1	bed2gff	feature	1048201	1050799	0	+	.	group3;
    chr1	bed2gff	feature	1264401	1266799	0	+	.	group4;
    chr1	bed2gff	feature	2008801	2011399	0	+	.	group5;
    chr1	bed2gff	feature	2185201	2188399	0	+	.	group6;
    chr1	bed2gff	feature	2577201	2578999	0	+	.	group7;
    chr1	bed2gff	feature	2786201	2789199	0	+	.	group8;
    chr1	bed2gff	feature	2923401	2926199	0	+	.	group9;
    chr1	bed2gff	feature	3256401	3264399	0	+	.	group10;
    Thanks for helping me out!

  • #2
    Please have a look at the GFF format description (e.g. here). You need to rephrase the last column to ID=group1; ID=group2; ...

    Cheers,
    Michael

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