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Report generated at 2020-05-28 13:15:12

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	63728292	137672758
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	63390988	135411543
Mapped(QC-failed)	0	0
% Mapped	99.4700	98.3600
Paired	63728292	137672758
Paired(QC-failed)	0	0
Read1	31864146	68836379
Read1(QC-failed)	0	0
Read2	31864146	68836379
Read2(QC-failed)	0	0
Properly Paired	63125333	130358651
Properly Paired(QC-failed)	0	0
% Properly Paired	99.0500	94.6900
With itself	63193885	134702187
With itself(QC-failed)	0	0
Singletons	197103	709356
Singletons(QC-failed)	0	0
% Singleton	0.3100	0.5200
Diff. Chroms	16258	242628
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	29679962	57571020
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	841829	552898
Paired Opt. Dupes	982	2174
% Dupes/100	0.0284	0.0096

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	29657596	57401418
Distinct Read Pairs	28817292	56929860
One Read Pair	28116011	56475460
Two Read Pairs	607895	445030
NRF = Distinct/Total	0.9717	0.9918
PBC1 = OnePair/Distinct	0.9757	0.9920
PBC2 = OnePair/TwoPair	46.2514	126.9026

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	57676266	114036244
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	57676266	114036244
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	57676266	114036244
Paired(QC-failed)	0	0
Read1	28838133	57018122
Read1(QC-failed)	0	0
Read2	28838133	57018122
Read2(QC-failed)	0	0
Properly Paired	57676266	114036244
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	57676266	114036244
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	98971
Np	0
N optimal	98971
N conservative	98971
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.	160
Corr. Est. Fragment Len.	0.2934
Phantom Peak	40
Corr. Phantom Peak	0.2241
Argmin. Corr.	1500
Min. Corr.	0.1690
NSC	1.7363
RSC	2.2571

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	0.6718

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	0.0812
AUC	0.4947
CHANCE divergence	0.2658
Elbow Point	0.0000
JS Distance	0.8864
Synthetic AUC	0.5037
Synthetic Elbow Point	0.5289
Synthetic JS Distance	0.6324