Pre-processing for diffHiC
1
0
Entering edit mode
@james-w-macdonald-5106
Last seen 1 day ago
United States

After aligning data using the presplit_map.py Python script, the next recommended step is to use Picard FixMateInformation and MarkDuplicates. However, using the current version of Picard I get

Exception in thread "main" htsjdk.samtools.SAMFormatException: SAM validation error: ERROR: Record 1, Read name K00280:111:HWNMLBBXX:7:1101:1174:14643, Mapped mate should have mate reference name

Which is hogwash:

$ samtools view 1-1.bam | head -n 4 | cut -f 1-2
K00280:111:HWNMLBBXX:7:1101:1174:14643    81
K00280:111:HWNMLBBXX:7:1101:1174:14643    145
K00280:111:HWNMLBBXX:7:1101:1174:14713    2049
K00280:111:HWNMLBBXX:7:1101:1174:14713    81

Anyway, is there some trickeration that needs to be done (other than using VALIDATION_STRINGENCY=DONTBOTHERMEABOUTFAKESTUFF)?

diffHiC • 886 views
ADD COMMENT
0
Entering edit mode
Aaron Lun ★ 28k
@alun
Last seen 7 hours ago
The city by the bay

Looking at https://github.com/LTLA/diffHicUsersGuide/blob/master/sra2bam.sh indicates that I did indeed set VALIDATION_STRINGENCY=SILENT. I recall that Picard would complain about everything and anything (especially prior to fixing the mate information) so you end up having to set this flag all the time.

ADD COMMENT
0
Entering edit mode

Thanks Aaron. SILENT seems to be the hot ticket. LENIENT, not so much.

ADD REPLY

Login before adding your answer.

Traffic: 318 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6