Untitled

No description

Report generated at 2020-05-14 16:27:22

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	95601862	152308104
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	91760373	148017607
Mapped(QC-failed)	0	0
% Mapped	95.9800	97.1800
Paired	95601862	152308104
Paired(QC-failed)	0	0
Read1	47800931	76154052
Read1(QC-failed)	0	0
Read2	47800931	76154052
Read2(QC-failed)	0	0
Properly Paired	90466260	145664311
Properly Paired(QC-failed)	0	0
% Properly Paired	94.6300	95.6400
With itself	91331636	147236867
With itself(QC-failed)	0	0
Singletons	428737	780740
Singletons(QC-failed)	0	0
% Singleton	0.4500	0.5100
Diff. Chroms	536390	863963
Diff. Chroms (QC-failed)	0	0

Marking duplicates (filtered BAM)

Filtered out (samtools view -F 1804):

read unmapped (0x4)
mate unmapped (0x8)
not primary alignment (0x100)
read fails platform/vendor quality checks (0x200)
read is PCR or optical duplicate (0x400)

	rep1	ctl1
Unpaired Reads	0	0
Paired Reads	41219414	63227368
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	149552	220283
Paired Opt. Dupes	1078	1066
% Dupes/100	0.0036	0.0035

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	41216497	63148791
Distinct Read Pairs	41066958	62930074
One Read Pair	40917885	62712523
Two Read Pairs	148613	216455
NRF = Distinct/Total	0.9964	0.9965
PBC1 = OnePair/Distinct	0.9964	0.9965
PBC2 = OnePair/TwoPair	275.3318	289.7255

Mitochondrial reads are filtered out.

NRF (non redundant fraction)
PBC1 (PCR Bottleneck coefficient 1)
PBC2 (PCR Bottleneck coefficient 2)
PBC1 is the primary measure. Provisionally

0-0.5 is severe bottlenecking
0.5-0.8 is moderate bottlenecking
0.8-0.9 is mild bottlenecking
0.9-1.0 is no bottlenecking

Flagstat (filtered/deduped BAM)

Filtered and duplicates removed

	rep1	ctl1
Total	82139724	126014170
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	82139724	126014170
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	82139724	126014170
Paired(QC-failed)	0	0
Read1	41069862	63007085
Read1(QC-failed)	0	0
Read2	41069862	63007085
Read2(QC-failed)	0	0
Properly Paired	82139724	126014170
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	82139724	126014170
With itself(QC-failed)	0	0
Singletons	0	0
Singletons(QC-failed)	0	0
% Singleton	0.0000	0.0000
Diff. Chroms	0	0
Diff. Chroms (QC-failed)	0	0

Peak calling

Reproducibility QC and peak detection statistics

The number of peaks is capped at 300K for peak-caller MACS2

	overlap
Nt	0
N1	210474
Np	0
N optimal	210474
N conservative	210474
Optimal Set	rep1-pr
Conservative Set	rep1-pr
Rescue Ratio	0.0000
Self Consistency Ratio	1.0000
Reproducibility	pass

Overlapping peaks

N1: Replicate 1 self-consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
N2: Replicate 2 self-consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
Nt: True Replicate consisten overlapping peaks (comparing true replicates Rep1 vs Rep2 )
Np: Pooled-pseudoreplicate consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling pooled reads from Rep1 and Rep2 )
Self-consistency Ratio: max(N1,N2) / min (N1,N2)
Rescue Ratio: max(Np,Nt) / min (Np,Nt)
Reproducibility Test: If Self-consistency Ratio >2 AND Rescue Ratio > 2, then 'Fail' else 'Pass'

Enrichment

Strand cross-correlation measures

Performed on subsampled reads (15M)

	rep1
Reads	15000000
Est. Fragment Len.	145
Corr. Est. Fragment Len.	0.1859
Phantom Peak	50
Corr. Phantom Peak	0.1923
Argmin. Corr.	1500
Min. Corr.	0.1772
NSC	1.0489
RSC	0.5734

NOTE1: For SE datasets, reads from replicates are randomly subsampled.
NOTE2: For PE datasets, the first end of each read-pair is selected and the reads are then randomly subsampled.

Normalized strand cross-correlation coefficient (NSC) = col9 in outFile
Relative strand cross-correlation coefficient (RSC) = col10 in outFile
Estimated fragment length = col3 in outFile, take the top value

Fraction of reads in overlapping peaks

	rep1-pr
Fraction of Reads in Peak	0.2950

ppr: Overlapping peaks comparing pooled pseudo replicates
rep1-pr: Overlapping peaks comparing pseudoreplicates from replicate 1
rep2-pr: Overlapping peaks comparing pseudoreplicates from replicate 2
repi-repj: Overlapping peaks comparing true replicates (rep i vs. rep j)

Other quality metrics

Fingerprint and Jensen-Shannon distance

	rep1
% genome enriched	0.2207
AUC	0.4955
CHANCE divergence	0.1176
Elbow Point	0.0000
JS Distance	0.6839
Synthetic AUC	0.4993
Synthetic Elbow Point	0.2161
Synthetic JS Distance	0.3734