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Report generated at 2021-02-05 11:56:33

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	55740606	120623008
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	55317617	118852959
Mapped(QC-failed)	0	0
% Mapped	99.2400	98.5300
Paired	55740606	120623008
Paired(QC-failed)	0	0
Read1	27870303	60311504
Read1(QC-failed)	0	0
Read2	27870303	60311504
Read2(QC-failed)	0	0
Properly Paired	54944137	116324482
Properly Paired(QC-failed)	0	0
% Properly Paired	98.5700	96.4400
With itself	55095510	118246914
With itself(QC-failed)	0	0
Singletons	222107	606045
Singletons(QC-failed)	0	0
% Singleton	0.4000	0.5000
Diff. Chroms	36941	142153
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	25228221	50494792
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	475796	383179
Paired Opt. Dupes	33921	18017
% Dupes/100	0.0189	0.0076

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	25188603	50475444
Distinct Read Pairs	24715093	50093864
One Read Pair	24283217	49738212
Two Read Pairs	398923	346479
NRF = Distinct/Total	0.9812	0.9924
PBC1 = OnePair/Distinct	0.9825	0.9929
PBC2 = OnePair/TwoPair	60.8719	143.5533

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	49504850	100223226
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	49504850	100223226
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	49504850	100223226
Paired(QC-failed)	0	0
Read1	24752425	50111613
Read1(QC-failed)	0	0
Read2	24752425	50111613
Read2(QC-failed)	0	0
Properly Paired	49504850	100223226
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	49504850	100223226
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	90155
Np	0
N optimal	90155
N conservative	90155
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.	155
Corr. Est. Fragment Len.	0.2751
Phantom Peak	55
Corr. Phantom Peak	0.2106
Argmin. Corr.	1500
Min. Corr.	0.1668
NSC	1.6488
RSC	2.4714

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	0.5671

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	0.1167
AUC	0.4940
CHANCE divergence	0.2078
Elbow Point	0.0000
JS Distance	0.8728
Synthetic AUC	0.5062
Synthetic Elbow Point	0.4668
Synthetic JS Distance	0.5691