I'm trying to bin 1x normalized ATAC-seq bigWig (generated by the bamCoverage function) with the multiBigwigSummary function in deepTools with the intention of clustering several ATAC-seq samples with deepTool's plotCorrelation function. My bamCoverage commands look like this:
bamCoverage -b input.bam -o output.SeqDepthNorm.bw -p "max" --effectiveGenomeSize 2805636331 --normalizeUsing RPGC -ignore chrX chrY chrM -e
This successfully generates the bigwig output files without any indication of a problem. However, when I then feed these bigwig files into multiBigwigSummary with the following command:
multiBigwigSummary bins --bwfiles first.SeqDepthNorm.bw \
second.SeqDepthNorm.bw \
third.SeqDepthNorm.bw \
fourth.SeqDepthNorm.bw \
fifth.SeqDepthNorm.bw \
sixth.SeqDepthNorm.bw \
seventh.SeqDepthNorm.bw \
--outFileName normalized_ATAC_peaks_bins.npz --binSize=1000 -p "max" -v \
--labels first second third fourth fifth sixth seventh
it throws the following error almost right away:
multiprocessing.pool.RemoteTraceback:
"""
Traceback (most recent call last):
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/multiprocessing/pool.py", line 119, in worker
result = (True, func(*args, **kwds))
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/multiprocessing/pool.py", line 44, in mapstar
return list(map(*args))
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/site-packages/deeptools/getScorePerBigWigBin.py", line 18, in countReadsInRegions_wrapper
return countFragmentsInRegions_worker(*args)
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/site-packages/deeptools/getScorePerBigWigBin.py", line 107, in countFragmentsInRegions_worker
score = bwh.stats(chrom, exon[0], exon[1])
RuntimeError: Invalid interval bounds!
"""
The above exception was the direct cause of the following exception:
Traceback (most recent call last):
File "/DCEG/Resources/Tools/python3/3.6.3-shared/bin/multiBigwigSummary", line 14, in <module>
main(args)
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/site-packages/deeptools/multiBigwigSummary.py", line 262, in main
allArgs=args)
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/site-packages/deeptools/getScorePerBigWigBin.py", line 268, in getScorePerBin
transcript_id_designator=transcript_id_designator)
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/site-packages/deeptools/mapReduce.py", line 142, in mapReduce
res = pool.map_async(func, TASKS).get(9999999)
File "/DCEG/Resources/Tools/python3/3.6.3-shared/lib/python3.6/multiprocessing/pool.py", line 644, in get
raise self._value
RuntimeError: Invalid interval bounds!
What's causing this problem? Is there a workaround? Any help ironing this out would be greatly appreciated!