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Report generated at 2021-03-18 16:35:40

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	19809550	140350086
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	16126085	136223961
Mapped(QC-failed)	0	0
% Mapped	81.4100	97.0600
Paired	19809550	140350086
Paired(QC-failed)	0	0
Read1	9904775	70175043
Read1(QC-failed)	0	0
Read2	9904775	70175043
Read2(QC-failed)	0	0
Properly Paired	15801473	120532551
Properly Paired(QC-failed)	0	0
% Properly Paired	79.7700	85.8800
With itself	16043570	134681268
With itself(QC-failed)	0	0
Singletons	82515	1542693
Singletons(QC-failed)	0	0
% Singleton	0.4200	1.1000
Diff. Chroms	92850	9773665
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	7302336	53789939
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	492243	594512
Paired Opt. Dupes	1664	1824
% Dupes/100	0.0674	0.0111

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	7301663	53785343
Distinct Read Pairs	6809459	53191094
One Read Pair	6370917	52633239
Two Read Pairs	404787	539980
NRF = Distinct/Total	0.9326	0.9890
PBC1 = OnePair/Distinct	0.9356	0.9895
PBC2 = OnePair/TwoPair	15.7389	97.4726

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	13620186	106390854
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	13620186	106390854
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	13620186	106390854
Paired(QC-failed)	0	0
Read1	6810093	53195427
Read1(QC-failed)	0	0
Read2	6810093	53195427
Read2(QC-failed)	0	0
Properly Paired	13620186	106390854
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	13620186	106390854
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	67249
Np	0
N optimal	67249
N conservative	67249
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 (8M)

	rep1
Reads	8106382
Est. Fragment Len.	150
Corr. Est. Fragment Len.	0.1894
Phantom Peak	55
Corr. Phantom Peak	0.1343
Argmin. Corr.	1500
Min. Corr.	0.0967
NSC	1.9584
RSC	2.4654

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	0.4569

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	0.0973
AUC	0.4890
CHANCE divergence	0.4984
Elbow Point	0.0000
JS Distance	0.8076
Synthetic AUC	0.5078
Synthetic Elbow Point	0.3969
Synthetic JS Distance	0.4841