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Report generated at 2020-07-10 17:14:04

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	103571170	112771886
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	98059137	109873578
Mapped(QC-failed)	0	0
% Mapped	94.6800	97.4300
Paired	103571170	112771886
Paired(QC-failed)	0	0
Read1	51785585	56385943
Read1(QC-failed)	0	0
Read2	51785585	56385943
Read2(QC-failed)	0	0
Properly Paired	76473680	84952591
Properly Paired(QC-failed)	0	0
% Properly Paired	73.8400	75.3300
With itself	95559095	108164942
With itself(QC-failed)	0	0
Singletons	2500042	1708636
Singletons(QC-failed)	0	0
% Singleton	2.4100	1.5200
Diff. Chroms	13369455	19230639
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	27637932	37110419
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	926313	616930
Paired Opt. Dupes	1737	2436
% Dupes/100	0.0335	0.0166

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	27605219	37079658
Distinct Read Pairs	26683080	36465928
One Read Pair	25889518	35868516
Two Read Pairs	753065	585863
NRF = Distinct/Total	0.9666	0.9834
PBC1 = OnePair/Distinct	0.9703	0.9836
PBC2 = OnePair/TwoPair	34.3789	61.2234

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	53423238	72986978
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	53423238	72986978
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	53423238	72986978
Paired(QC-failed)	0	0
Read1	26711619	36493489
Read1(QC-failed)	0	0
Read2	26711619	36493489
Read2(QC-failed)	0	0
Properly Paired	53423238	72986978
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	53423238	72986978
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	184680
Np	0
N optimal	184680
N conservative	184680
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.	0.1954
Phantom Peak	50
Corr. Phantom Peak	0.2199
Argmin. Corr.	1500
Min. Corr.	0.1809
NSC	1.0801
RSC	0.3712

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	0.2464

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	0.2163
AUC	0.4944
CHANCE divergence	0.1427
Elbow Point	0.0000
JS Distance	0.6799
Synthetic AUC	0.5002
Synthetic Elbow Point	0.1939
Synthetic JS Distance	0.3669