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Report generated at 2020-06-08 02:03:41

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	235817820	92143016
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	229674000	82943830
Mapped(QC-failed)	0	0
% Mapped	97.3900	90.0200
Paired	235817820	92143016
Paired(QC-failed)	0	0
Read1	117908910	46071508
Read1(QC-failed)	0	0
Read2	117908910	46071508
Read2(QC-failed)	0	0
Properly Paired	227530091	75643794
Properly Paired(QC-failed)	0	0
% Properly Paired	96.4900	82.0900
With itself	228609726	81703578
With itself(QC-failed)	0	0
Singletons	1064274	1240252
Singletons(QC-failed)	0	0
% Singleton	0.4500	1.3500
Diff. Chroms	264196	199484
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	101309983	33771535
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	14248223	1015840
Paired Opt. Dupes	22089	11140
% Dupes/100	0.1406	0.0301

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	101305313	33756367
Distinct Read Pairs	87057941	32741033
One Read Pair	75183146	31771390
Two Read Pairs	9885650	934950
NRF = Distinct/Total	0.8594	0.9699
PBC1 = OnePair/Distinct	0.8636	0.9704
PBC2 = OnePair/TwoPair	7.6053	33.9819

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	174123520	65511390
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	174123520	65511390
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	174123520	65511390
Paired(QC-failed)	0	0
Read1	87061760	32755695
Read1(QC-failed)	0	0
Read2	87061760	32755695
Read2(QC-failed)	0	0
Properly Paired	174123520	65511390
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	174123520	65511390
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	83842
Np	0
N optimal	83842
N conservative	83842
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.	140
Corr. Est. Fragment Len.	0.1724
Phantom Peak	50
Corr. Phantom Peak	0.1826
Argmin. Corr.	1500
Min. Corr.	0.1685
NSC	1.0229
RSC	0.2740

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	0.0524

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	0.3320
AUC	0.4969
CHANCE divergence	0.0964
Elbow Point	0.0000
JS Distance	0.5439
Synthetic AUC	0.5032
Synthetic Elbow Point	0.0761
Synthetic JS Distance	0.2021