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Report generated at 2020-05-13 10:08:44

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	66354766	61233540
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	55932889	58236787
Mapped(QC-failed)	0	0
% Mapped	84.2900	95.1100
Paired	66354766	61233540
Paired(QC-failed)	0	0
Read1	33177383	30616770
Read1(QC-failed)	0	0
Read2	33177383	30616770
Read2(QC-failed)	0	0
Properly Paired	53470062	50526323
Properly Paired(QC-failed)	0	0
% Properly Paired	80.5800	82.5100
With itself	54501825	56771071
With itself(QC-failed)	0	0
Singletons	1431064	1465716
Singletons(QC-failed)	0	0
% Singleton	2.1600	2.3900
Diff. Chroms	761541	4278577
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	24108964	21455327
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	472424	305205
Paired Opt. Dupes	2199	1806
% Dupes/100	0.0196	0.0142

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	24097225	21407515
Distinct Read Pairs	23625093	21105831
One Read Pair	23160816	20807859
Two Read Pairs	456521	294314
NRF = Distinct/Total	0.9804	0.9859
PBC1 = OnePair/Distinct	0.9803	0.9859
PBC2 = OnePair/TwoPair	50.7333	70.6995

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	47273080	42300244
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	47273080	42300244
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	47273080	42300244
Paired(QC-failed)	0	0
Read1	23636540	21150122
Read1(QC-failed)	0	0
Read2	23636540	21150122
Read2(QC-failed)	0	0
Properly Paired	47273080	42300244
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	47273080	42300244
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	73513
Np	0
N optimal	73513
N conservative	73513
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.	255
Corr. Est. Fragment Len.	0.2172
Phantom Peak	50
Corr. Phantom Peak	0.2111
Argmin. Corr.	1500
Min. Corr.	0.1848
NSC	1.1752
RSC	1.2344

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	0.2949

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	0.1434
AUC	0.4940
CHANCE divergence	0.3352
Elbow Point	0.0000
JS Distance	0.6946
Synthetic AUC	0.4946
Synthetic Elbow Point	0.2157
Synthetic JS Distance	0.4339