LieberInstitute / RNAseq-pipeline

Original version of the RNA-seq pipeline implemented in SPEAQeasy at https://github.com/LieberInstitute/SPEAQeasy.
http://research.libd.org/SPEAQeasy/
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Error in creating the BigWig files when the data is unstranded #26

Closed lcolladotor closed 7 years ago

lcolladotor commented 7 years ago

http://rseqc.sourceforge.net/#bam2wig-py specifies that -d STRAND_RULE has by default the value "none". Turns out that using "none" leads to log files like this one:

**** Job starts ****
Mon Feb 20 10:19:16 EST 2017
Calcualte wigsum ... 
No alignments for GL383567.1. skipped
Processing KI270509.1 ...
Processing KI270302.1 ...
No alignments for KI270787.1. skipped
Processing KI270374.1 ...
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Processing KI270424.1 ...
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Processing KI270322.1 ...
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Processing KI270448.1 ...
Processing KI270438.1 ...
Processing GL000008.2 ...
No alignments for KI270888.1. skipped
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Processing KI270378.1 ...
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Processing KI270466.1 ...
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Processing KI270521.1 ...
Processing KI270412.1 ...
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Processing KI270372.1 ...
Processing GL000208.1 ...
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Processing KI270376.1 ...
Processing KI270320.1 ...
Processing KI270442.1 ...
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Processing KI270720.1 ...
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Processing KI270722.1 ...
Processing KI270414.1 ...
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Processing KI270316.1 ...
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Processing KI270724.1 ...
Processing KI270330.1 ...
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Processing KI270338.1 ...
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Processing KI270312.1 ...
No alignments for JH159148.1. skipped
Processing KI270726.1 ...
Processing KI270336.1 ...
Processing KI270728.1 ...
Processing KI270420.1 ...
No alignments for KI270884.1. skipped
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Processing KI270706.1 ...
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Processing KI270334.1 ...
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Processing KI270746.1 ...
Processing chrY ...
Processing chrX ...
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Processing chrM ...
Processing KI270732.1 ...
Processing GL000194.1 ...
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Processing KI270730.1 ...
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Processing KI270388.1 ...
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Processing KI270736.1 ...
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Processing KI270582.1 ...
No alignments for GL000209.2. skipped
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Processing chr7 ...
Processing KI270742.1 ...
Processing chr5 ...
Processing chr4 ...
Processing chr3 ...
Processing chr2 ...
Processing chr1 ...
Processing KI270584.1 ...
Processing KI270734.1 ...
Processing chr9 ...
Processing chr8 ...
No alignments for KI270874.1. skipped
Processing KI270740.1 ...
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Processing GL000219.1 ...
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Processing chr13 ...
Processing chr12 ...
Processing chr11 ...
Processing chr10 ...
Processing chr17 ...
Processing chr16 ...
Processing chr15 ...
Processing chr14 ...
Processing chr19 ...
Processing chr18 ...
Processing KI270718.1 ...
No alignments for KI270808.1. skipped
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Processing GL000205.2 ...
Processing GL000213.1 ...
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Processing KI270580.1 ...
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Processing KI270750.1 ...
Processing KI270382.1 ...
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Processing KI270708.1 ...
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Processing KI270588.1 ...
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Processing KI270590.1 ...
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Processing KI270386.1 ...
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Processing KI270748.1 ...
No alignments for GL383546.1. skipped
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Processing KI270384.1 ...
Processing KI270754.1 ...
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Processing GL000221.1 ...
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Processing KI270756.1 ...
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Processing KI270738.1 ...
Processing KI270744.1 ...
Processing KI270714.1 ...
No alignments for GL383540.1. skipped
Processing KI270544.1 ...
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Processing KI270394.1 ...
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Processing KI270528.1 ...
Processing GL000225.1 ...
Processing KI270396.1 ...
Processing chr6 ...
No alignments for GL383528.1. skipped
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Processing KI270390.1 ...
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Processing KI270392.1 ...
Processing KI270516.1 ...
Processing KI270712.1 ...
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Processing KI270538.1 ...
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Processing KI270508.1 ...
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Processing KI270375.1 ...
Processing KI270425.1 ...
Processing KI270752.1 ...
Processing KI270379.1 ...
No alignments for GL383576.1. skipped
Processing KI270522.1 ...
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Processing KI270423.1 ...
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Processing GL000218.1 ...
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Processing KI270435.1 ...
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Processing KI270373.1 ...
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Processing KI270371.1 ...
Processing KI270465.1 ...
Processing KI270710.1 ...
No alignments for KI270897.1. skipped
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Processing KI270315.1 ...
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Processing KI270363.1 ...
Processing KI270518.1 ...
Processing KI270317.1 ...
Processing KI270548.1 ...
No alignments for GL383532.1. skipped
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Processing KI270510.1 ...
Processing KI270311.1 ...
Processing KI270419.1 ...
No alignments for KI270812.1. skipped
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Processing KI270303.1 ...
Processing KI270429.1 ...
Processing KI270337.1 ...
Processing KI270335.1 ...
No alignments for KI270918.1. skipped
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Processing KI270530.1 ...
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Processing KI270305.1 ...
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Processing KI270739.1 ...
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Processing KI270733.1 ...
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Processing GL000195.1 ...
Processing KI270731.1 ...
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Processing KI270737.1 ...
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Processing KI270729.1 ...
Processing KI270411.1 ...
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Processing KI270417.1 ...
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Processing KI270721.1 ...
No alignments for KI270805.1. skipped
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No alignments for KI270920.1. skipped
Processing KI270329.1 ...
Processing KI270723.1 ...
Processing KI270735.1 ...
Processing KI270725.1 ...
No alignments for GL383582.2. skipped
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Processing KI270713.1 ...
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Processing KI270719.1 ...
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Processing KI270727.1 ...
No alignments for KI270801.1. skipped
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Processing GL000214.1 ...
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Processing KI270707.1 ...
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Processing GL000216.2 ...
Processing KI270711.1 ...
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Processing KI270591.1 ...
Processing KI270747.1 ...
No alignments for KI270775.1. skipped
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Processing KI270304.1 ...
Processing KI270755.1 ...
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Processing KI270757.1 ...
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Processing KI270583.1 ...
No alignments for KI270765.1. skipped
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Processing KI270743.1 ...
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Processing KI270715.1 ...
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Processing GL000009.2 ...
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Processing GL000220.1 ...
Processing KI270709.1 ...
Processing KI270383.1 ...
Processing KI270391.1 ...
No alignments for GL383555.2. skipped
Processing KI270517.1 ...
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Processing KI270393.1 ...
Processing KI270589.1 ...
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No alignments for GL000254.2. skipped
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Processing KI270381.1 ...
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Processing KI270387.1 ...
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Processing KI270385.1 ...
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Processing KI270507.1 ...
Processing KI270753.1 ...
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Processing KI270749.1 ...
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Processing KI270366.1 ...
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Processing KI270745.1 ...
No alignments for GL383573.1. skipped
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Processing KI270468.1 ...
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Processing KI270529.1 ...
Processing GL000224.1 ...
No alignments for GL383579.2. skipped
Processing KI270422.1 ...
Processing GL000226.1 ...
Processing KI270362.1 ...
Processing chr22 ...
Processing chr20 ...
Processing chr21 ...
Processing KI270340.1 ...
Processing KI270519.1 ...
Processing KI270364.1 ...
No alignments for GL383521.1. skipped
No alignments for GL383533.1. skipped
Processing KI270418.1 ...
No alignments for GL383527.1. skipped
Processing KI270511.1 ...

total wigsum is:8063646795.0

Unknown value of option :'strand_rule' none
Skip multi-hits:False
rm: cannot remove `/dcl01/lieber/ajaffe/lab/brainseq_phase2/degradation/DLPFC_polyA/Coverage/DLPFC_Br2020_C1_poly*.wig': No such file or directory
**** Job ends ****
Mon Feb 20 10:23:36 EST 2017

I'm checking what's the correct way to specify that the data is unstranded. It might be -d "" or simply without using -d.

lcolladotor commented 7 years ago

Using -d "" leads to:

Unknown value of option :'strand_rule'
lcolladotor commented 7 years ago

If you run the tests, you'll note that stranded single-end data fails. Since that type of data is not supported by the pipeline yet, it should be fine. Anyhow, just something to take note of @emilyburke