featureCounts: No effect of setting -d and -D?
1
0
Entering edit mode
@darioberaldi-6991
Last seen 2.1 years ago
United Kingdom

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)
subread featureCounts • 1.0k views
ADD COMMENT
2
Entering edit mode
ATpoint ★ 4.6k
@atpoint-13662
Last seen 14 hours ago
Germany

I think you additionally need -P (capital P) to trigger that the D/d options are enforced. Should be implied when using D/d but apparently it isn't.

ADD COMMENT
0
Entering edit mode

Yes, well spotted. Also -B is required when using -P but at least you get a meaningful error if you omit -B.

ADD REPLY

Login before adding your answer.

Traffic: 695 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