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Old 11-20-2013, 05:52 PM   #1
norafar
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Location: New York, USA

Join Date: May 2013
Posts: 5
Default HTSeq issue with stdout/stdin

Hello All,

I like to apply HTSeq-count on a .bam file, but if I directly pipe the output of samtools to htseq it gives me an error. However, if I first write the sorted file in a temporary file and then apply samtools-view and htseq-count on this temp file, there is no error!
Do you know what might be causing this problem?

Case (1): [generates error]

$ samtools sort -on -m 300000000 accepted_hits.bam | samtools view -h - | htseq-count -m union - hg19.gtf > output.count

Error occured when reading first line of sam file.
[Exception type: StopIteration, raised in count.py:83]


Case (2): [no error!]
$ samtools sort -n -m 300000000 accepted_hits.bam temp-sorted.bam
$ samtools view -h temp-sorted.bam | htseq-count -m union - hg19.gtf > output.count

Thank You,
NF
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Old 11-27-2013, 11:55 PM   #2
Simon Anders
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Location: Heidelberg, Germany

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Default

'samtools sort' does not write to stdout, so you cannot use piping.
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Old 11-30-2013, 04:22 PM   #3
arm55
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Default

norafar, include the - file marker as output on the sort and it should work.

samtools sort -on -m 300000000 accepted_hits.bam - | samtools view -h - | htseq-count -m union - hg19.gtf > output.count

Last edited by arm55; 11-30-2013 at 04:25 PM. Reason: Better answer
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htseq, htseq count, rna seq

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