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Report generated at 2022-01-05 15:45:22

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	53805012	146003154
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	53074143	143950255
Mapped(QC-failed)	0	0
% Mapped	98.6400	98.5900
Paired	53805012	146003154
Paired(QC-failed)	0	0
Read1	26902506	73001577
Read1(QC-failed)	0	0
Read2	26902506	73001577
Read2(QC-failed)	0	0
Properly Paired	52495085	139355986
Properly Paired(QC-failed)	0	0
% Properly Paired	97.5700	95.4500
With itself	52802636	143134940
With itself(QC-failed)	0	0
Singletons	271507	815315
Singletons(QC-failed)	0	0
% Singleton	0.5000	0.5600
Diff. Chroms	78827	200467
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	23147561	61294235
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	175887	434896
Paired Opt. Dupes	12888	3707
% Dupes/100	0.0076	0.0071

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	23060795	61180783
Distinct Read Pairs	22899628	60780225
One Read Pair	22742158	60396899
Two Read Pairs	155020	377062
NRF = Distinct/Total	0.9930	0.9935
PBC1 = OnePair/Distinct	0.9931	0.9937
PBC2 = OnePair/TwoPair	146.7047	160.1776

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	45943348	121718678
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	45943348	121718678
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	45943348	121718678
Paired(QC-failed)	0	0
Read1	22971674	60859339
Read1(QC-failed)	0	0
Read2	22971674	60859339
Read2(QC-failed)	0	0
Properly Paired	45943348	121718678
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	45943348	121718678
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	49836
Np	0
N optimal	49836
N conservative	49836
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.	-5
Corr. Est. Fragment Len.	0.1764
Phantom Peak	50
Corr. Phantom Peak	0.1820
Argmin. Corr.	1500
Min. Corr.	0.1686
NSC	1.0464
RSC	0.5806

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	0.1101

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	0.2867
AUC	0.4940
CHANCE divergence	0.1127
Elbow Point	0.0000
JS Distance	0.5796
Synthetic AUC	0.5016
Synthetic Elbow Point	0.1421
Synthetic JS Distance	0.2674