Closed wrshoemaker closed 6 years ago
Hi Will,
Did you do this test using the same SAM file that you had used previously? From the error and the output it looks like there is an issue with the mapping - the genome you are using has zero coverage.
Chris
On Sep 21, 2017, at 5:50 PM, Will Shoemaker notifications@github.com wrote:
Hello,
I really liked iRep and have used it before. I re-installed it a few weeks ago got some weird results that I think might be caused by numpy code in the iRep package.
When I run iRep on my sorted sam file I get the following output and the following numpy warnings:
~/iRep/lib/python3.6/site-packages/numpy/lib/function_base.py:1110: RuntimeWarning: Mean of empty slice. avg = a.mean(axis)
and
~/iRep/lib/python3.6/site-packages/numpy/core/_methods.py:80: RuntimeWarning: invalid value encountered in double_scalars ret = ret.dtype.type(ret / rcount)
I think this might have something to do with the sliding window calculation. Have you seen something similar and do you know how to work around it?
— You are receiving this because you are subscribed to this thread. Reply to this email directly, view it on GitHub, or mute the thread.
Hi Chris,
I'm not using the same SAM file, but this SAM file has an average coverage of ~500. I also sorted it using the samtools sort
command and checked to make sure that the reference sequence names in the SAM file match the sequence names in the FASTA file.
I can send a portion of the SAM file if that would be useful. The entire file is ~ 7 gb.
Hi Will,
Interesting. Can you please confirm that the read mapping was done in paired-end mode (e.g. using -1 and -2 with Bowtie2)?
Other than that, I'm not sure what the issue could be. It would be helpful if you could send the first few lines of your SAM mapping file.
Sorry for the inconvenience.
Chris
On Sep 22, 2017, at 11:09 AM, Will Shoemaker notifications@github.com wrote:
I can send a portion of the SAM file if that would be useful. The entire file is ~ 7 gb.
— You are receiving this because you commented. Reply to this email directly, view it on GitHub, or mute the thread.
On Sep 22, 2017, at 10:06 AM, Will Shoemaker notifications@github.com wrote:
Hi Chris,
I'm not using the same SAM file, but this SAM file has an average coverage of ~500. I also sorted it using the samtools sort command and checked to make sure that the reference sequence names in the SAM file match the sequence names in the FASTA file.
— You are receiving this because you commented. Reply to this email directly, view it on GitHub, or mute the thread.
Hi Chris,
I mapped the reads to the reference using breseq
, which treats the reads of single-end. I wasn't aware that the reads needed to be paired-end. I'll re-map the reads to the reference, re-run iRep
, and see if there's still a problem.
Thanks for your help, Will
Hi Will,
Hopefully that will fix it for you.
iRep does look at paired read mappings to make sure that both reads in each pair are accurately mapped to the genome. This makes it much less likely to have off target read mappings that could inflate coverage values over some parts of the gnome, and skew results.
Chris
On Sep 22, 2017, at 2:31 PM, Will Shoemaker notifications@github.com wrote:
Hi Chris,
I mapped the reads to the reference using breseq, which treats the reads of single-end. I wasn't aware that the reads needed to be paired-end. I'll re-map the reads to the reference, re-run iRep, and see if there's still a problem.
Thanks for your help, Will
— You are receiving this because you commented. Reply to this email directly, view it on GitHub, or mute the thread.
Thanks Chris,
That solved the problem. Glad to know there's that extra step in iRep. I'll go ahead and close this "issue".
Hello,
I really liked iRep and have used it before. I re-installed it a few weeks ago got some weird results that I think might be caused by numpy code in the iRep package.
When I run iRep on my sorted sam file I get the following output and the following numpy warnings:
~/iRep/lib/python3.6/site-packages/numpy/lib/function_base.py:1110: RuntimeWarning: Mean of empty slice. avg = a.mean(axis)
and
~/iRep/lib/python3.6/site-packages/numpy/core/_methods.py:80: RuntimeWarning: invalid value encountered in double_scalars ret = ret.dtype.type(ret / rcount)
I think this might have something to do with the sliding window calculation. Have you seen something similar and do you know how to work around it?