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Report generated at 2020-05-18 21:31:05

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	110538002	168466658
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	105109430	164624745
Mapped(QC-failed)	0	0
% Mapped	95.0900	97.7200
Paired	110538002	168466658
Paired(QC-failed)	0	0
Read1	55269001	84233329
Read1(QC-failed)	0	0
Read2	55269001	84233329
Read2(QC-failed)	0	0
Properly Paired	103346698	162184918
Properly Paired(QC-failed)	0	0
% Properly Paired	93.4900	96.2700
With itself	104522762	163825228
With itself(QC-failed)	0	0
Singletons	586668	799517
Singletons(QC-failed)	0	0
% Singleton	0.5300	0.4700
Diff. Chroms	671008	775185
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	45807959	70866978
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	255398	312582
Paired Opt. Dupes	1361	1250
% Dupes/100	0.0056	0.0044

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	45803382	70788047
Distinct Read Pairs	45548020	70477116
One Read Pair	45294130	70167781
Two Read Pairs	252429	307786
NRF = Distinct/Total	0.9944	0.9956
PBC1 = OnePair/Distinct	0.9944	0.9956
PBC2 = OnePair/TwoPair	179.4331	227.9759

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	91105122	141108792
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	91105122	141108792
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	91105122	141108792
Paired(QC-failed)	0	0
Read1	45552561	70554396
Read1(QC-failed)	0	0
Read2	45552561	70554396
Read2(QC-failed)	0	0
Properly Paired	91105122	141108792
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	91105122	141108792
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	211394
Np	0
N optimal	211394
N conservative	211394
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.	145
Corr. Est. Fragment Len.	0.1912
Phantom Peak	50
Corr. Phantom Peak	0.2029
Argmin. Corr.	1500
Min. Corr.	0.1822
NSC	1.0494
RSC	0.4339

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	0.3649

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	0.2081
AUC	0.4957
CHANCE divergence	0.1145
Elbow Point	0.0000
JS Distance	0.7240
Synthetic AUC	0.5019
Synthetic Elbow Point	0.2546
Synthetic JS Distance	0.3993