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Report generated at 2021-02-06 20:25:42

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	87615912	148269018
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	86462773	143500908
Mapped(QC-failed)	0	0
% Mapped	98.6800	96.7800
Paired	87615912	148269018
Paired(QC-failed)	0	0
Read1	43807956	74134509
Read1(QC-failed)	0	0
Read2	43807956	74134509
Read2(QC-failed)	0	0
Properly Paired	85459538	140225987
Properly Paired(QC-failed)	0	0
% Properly Paired	97.5400	94.5800
With itself	86102464	141622671
With itself(QC-failed)	0	0
Singletons	360309	1878237
Singletons(QC-failed)	0	0
% Singleton	0.4100	1.2700
Diff. Chroms	354394	652729
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	38112353	59090669
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	367876	523701
Paired Opt. Dupes	3679	14446
% Dupes/100	0.0097	0.0089

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	38094862	57674075
Distinct Read Pairs	37727199	57472830
One Read Pair	37362863	57272993
Two Read Pairs	361040	198503
NRF = Distinct/Total	0.9903	0.9965
PBC1 = OnePair/Distinct	0.9903	0.9965
PBC2 = OnePair/TwoPair	103.4868	288.5246

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	75488954	117133936
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	75488954	117133936
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	75488954	117133936
Paired(QC-failed)	0	0
Read1	37744477	58566968
Read1(QC-failed)	0	0
Read2	37744477	58566968
Read2(QC-failed)	0	0
Properly Paired	75488954	117133936
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	75488954	117133936
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	100680
Np	0
N optimal	100680
N conservative	100680
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.	170
Corr. Est. Fragment Len.	0.2200
Phantom Peak	50
Corr. Phantom Peak	0.2197
Argmin. Corr.	1500
Min. Corr.	0.1883
NSC	1.1686
RSC	1.0092

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	0.3359

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	0.1943
AUC	0.4953
CHANCE divergence	0.1422
Elbow Point	0.0000
JS Distance	0.7179
Synthetic AUC	0.5059
Synthetic Elbow Point	0.3236
Synthetic JS Distance	0.4253