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Report generated at 2021-02-10 00:13:07

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	87160874	193594802
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	83614727	187969057
Mapped(QC-failed)	0	0
% Mapped	95.9300	97.0900
Paired	87160874	193594802
Paired(QC-failed)	0	0
Read1	43580437	96797401
Read1(QC-failed)	0	0
Read2	43580437	96797401
Read2(QC-failed)	0	0
Properly Paired	82576371	184287983
Properly Paired(QC-failed)	0	0
% Properly Paired	94.7400	95.1900
With itself	82964505	185784698
With itself(QC-failed)	0	0
Singletons	650222	2184359
Singletons(QC-failed)	0	0
% Singleton	0.7500	1.1300
Diff. Chroms	169594	610670
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	36763624	78211821
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	681662	539110
Paired Opt. Dupes	6617	19807
% Dupes/100	0.0185	0.0069

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	36687143	77646644
Distinct Read Pairs	36007733	77172156
One Read Pair	35339673	76701136
Two Read Pairs	656890	467668
NRF = Distinct/Total	0.9815	0.9939
PBC1 = OnePair/Distinct	0.9814	0.9939
PBC2 = OnePair/TwoPair	53.7985	164.0077

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	72163924	155345422
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	72163924	155345422
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	72163924	155345422
Paired(QC-failed)	0	0
Read1	36081962	77672711
Read1(QC-failed)	0	0
Read2	36081962	77672711
Read2(QC-failed)	0	0
Properly Paired	72163924	155345422
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	72163924	155345422
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	83059
Np	0
N optimal	83059
N conservative	83059
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.	150
Corr. Est. Fragment Len.	0.1855
Phantom Peak	50
Corr. Phantom Peak	0.1960
Argmin. Corr.	1500
Min. Corr.	0.1755
NSC	1.0572
RSC	0.4877

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	0.1677

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	0.2716
AUC	0.4952
CHANCE divergence	0.1042
Elbow Point	0.0000
JS Distance	0.6179
Synthetic AUC	0.4983
Synthetic Elbow Point	0.1867
Synthetic JS Distance	0.3028