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Report generated at 2020-05-23 10:38:54

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	176394838	145469574
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	166314878	141586568
Mapped(QC-failed)	0	0
% Mapped	94.2900	97.3300
Paired	176394838	145469574
Paired(QC-failed)	0	0
Read1	88197419	72734787
Read1(QC-failed)	0	0
Read2	88197419	72734787
Read2(QC-failed)	0	0
Properly Paired	163732280	139229482
Properly Paired(QC-failed)	0	0
% Properly Paired	92.8200	95.7100
With itself	165168945	140814697
With itself(QC-failed)	0	0
Singletons	1145933	771871
Singletons(QC-failed)	0	0
% Singleton	0.6500	0.5300
Diff. Chroms	615808	836459
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	70084103	60565195
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	518367	282279
Paired Opt. Dupes	1047	2180
% Dupes/100	0.0074	0.0047

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	70079700	60530190
Distinct Read Pairs	69561373	60248362
One Read Pair	69047698	59967875
Two Read Pairs	509093	279160
NRF = Distinct/Total	0.9926	0.9953
PBC1 = OnePair/Distinct	0.9926	0.9953
PBC2 = OnePair/TwoPair	135.6288	214.8154

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	139131472	120565832
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	139131472	120565832
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	139131472	120565832
Paired(QC-failed)	0	0
Read1	69565736	60282916
Read1(QC-failed)	0	0
Read2	69565736	60282916
Read2(QC-failed)	0	0
Properly Paired	139131472	120565832
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	139131472	120565832
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	99345
Np	0
N optimal	99345
N conservative	99345
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.	120
Corr. Est. Fragment Len.	0.1851
Phantom Peak	50
Corr. Phantom Peak	0.2067
Argmin. Corr.	1500
Min. Corr.	0.1774
NSC	1.0435
RSC	0.2627

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	0.0615

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	0.2924
AUC	0.4966
CHANCE divergence	0.0939
Elbow Point	0.0000
JS Distance	0.5772
Synthetic AUC	0.5035
Synthetic Elbow Point	0.0495
Synthetic JS Distance	0.2655