Entering edit mode
Using featureCounts v2.0.1, I don't see any effect in changing -d
and -D
options (minimum and maximum fragment/template length, respectively). My bam file has paired reads and I set the -p
option. Am I missing something?
Here's example output using silly values of -d 10 -D 10000
and -d 10000 -D 10
. No difference:
featureCounts -d 10 -D 10000 -a genrich/ATACseq/single/atac_merge.gff -o tmp.tsv -t peak -g locus_id -p bwa/Ap2oGFP1o.pb.bam
cat tmp.tsv.summary
Status bwa/Ap2oGFP1o.pb.bam
Assigned 3747979
Unassigned_Unmapped 0
Unassigned_Read_Type 0
Unassigned_Singleton 0
Unassigned_MappingQuality 0
Unassigned_Chimera 0
Unassigned_FragmentLength 0
Unassigned_Duplicate 0
Unassigned_MultiMapping 0
Unassigned_Secondary 0
Unassigned_NonSplit 0
Unassigned_NoFeatures 7721454
Unassigned_Overlapping_Length 0
Unassigned_Ambiguity 42450
featureCounts -d 10000 -D 10 -a genrich/ATACseq/single/atac_merge.gff -o tmp.tsv -t peak -g locus_id -p bwa/Ap2oGFP1o.pb.bam
cat tmp.tsv.summary
Status bwa/Ap2oGFP1o.pb.bam
Assigned 3747979
Unassigned_Unmapped 0
Unassigned_Read_Type 0
Unassigned_Singleton 0
Unassigned_MappingQuality 0
Unassigned_Chimera 0
Unassigned_FragmentLength 0
Unassigned_Duplicate 0
Unassigned_MultiMapping 0
Unassigned_Secondary 0
Unassigned_NonSplit 0
Unassigned_NoFeatures 7721454
Unassigned_Overlapping_Length 0
Unassigned_Ambiguity 42450
Summary of bam file:
samtools flagstat bwa/Ap2oGFP1o.pb.bam
23008511 + 0 in total (QC-passed reads + QC-failed reads)
23008511 + 0 primary
0 + 0 secondary
0 + 0 supplementary
1391376 + 0 duplicates
1391376 + 0 primary duplicates
23008511 + 0 mapped (100.00% : N/A)
23008511 + 0 primary mapped (100.00% : N/A)
23008511 + 0 paired in sequencing
11504546 + 0 read1
11503965 + 0 read2
23008511 + 0 properly paired (100.00% : N/A)
23008511 + 0 with itself and mate mapped
0 + 0 singletons (0.00% : N/A)
0 + 0 with mate mapped to a different chr
0 + 0 with mate mapped to a different chr (mapQ>=5)
Yes, well spotted. Also
-B
is required when using-P
but at least you get a meaningful error if you omit-B
.