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7 months ago
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I'm running rmats/4.1.2 to detect the splicing events.
python /software/rmats/4.1.2/rmats.py --gtf /reference_annotation/Homo_sapiens.GRCh38.86.gtf -t paired \
--b1 ko_1.txt --b2 wt_1.txt --od /Rmats_test \
--tmp /Rmats_test --libType fr-secondstrand --readLength 63 \
--nthread 4 --tstat 4 --variable-read-length --novelSS --allow-clipping
From the outcome file
USED: 49050150
NOT_PAIRED: 0
NOT_NH_1: 0
NOT_EXPECTED_CIGAR: 3190321
NOT_EXPECTED_READ_LENGTH: 0
NOT_EXPECTED_STRAND: 0
EXON_NOT_MATCHED_TO_ANNOTATION: 354260651
JUNCTION_NOT_MATCHED_TO_ANNOTATION: 53086428
CLIPPED: 0
TOTAL_FOR_BAM: 459587550
WT_1out.bam
Same report for the other file also. I have huge number of NOT MATCHED alignment between my BAM and GTF. I checked they are the same GFT file. Is there any other reason? Second one is NOT_PAIRED: 0 I did samtools stat
SN reads mapped and paired: 459587550 # paired-end technology bit set + both mates mapped
SN reads properly paired: 459587550 # proper-pair bit set
SN reads paired: 459587550 # paired-end technology bit set
SN inward oriented pairs: 229735745
SN outward oriented pairs: 58030
SN pairs with other orientation: 0
SN pairs on different chromosomes: 0
SN percentage of properly paired reads (%): 100.0
So i'm confused now. is there any qc i can do? any suggestion?
Thanks