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Report generated at 2019-10-13 06:49:41

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	86863022	87587868
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	82938549	85042791
Mapped(QC-failed)	0	0
% Mapped	95.4800	97.0900
Paired	86863022	87587868
Paired(QC-failed)	0	0
Read1	43431511	43793934
Read1(QC-failed)	0	0
Read2	43431511	43793934
Read2(QC-failed)	0	0
Properly Paired	81062990	83193525
Properly Paired(QC-failed)	0	0
% Properly Paired	93.3200	94.9800
With itself	81772327	83956805
With itself(QC-failed)	0	0
Singletons	1166222	1085986
Singletons(QC-failed)	0	0
% Singleton	1.3400	1.2400
Diff. Chroms	264219	333038
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	33242939	36699350
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	923285	221028
Paired Opt. Dupes	6978	15561
% Dupes/100	0.0278	0.0060

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	33242431	36659304
Distinct Read Pairs	32319153	36439024
One Read Pair	31442990	36223105
Two Read Pairs	846286	213134
NRF = Distinct/Total	0.9722	0.9940
PBC1 = OnePair/Distinct	0.9729	0.9941
PBC2 = OnePair/TwoPair	37.1541	169.9546

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	64639308	72956644
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	64639308	72956644
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	64639308	72956644
Paired(QC-failed)	0	0
Read1	32319654	36478322
Read1(QC-failed)	0	0
Read2	32319654	36478322
Read2(QC-failed)	0	0
Properly Paired	64639308	72956644
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	64639308	72956644
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	97938
Np	0
N optimal	97938
N conservative	97938
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.	125
Corr. Est. Fragment Len.	0.1805
Phantom Peak	50
Corr. Phantom Peak	0.2065
Argmin. Corr.	1500
Min. Corr.	0.1753
NSC	1.0300
RSC	0.1684

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	0.0717

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	0.2770
AUC	0.4949
CHANCE divergence	0.1175
Elbow Point	0.0000
JS Distance	0.5735
Synthetic AUC	0.5026
Synthetic Elbow Point	0.0942
Synthetic JS Distance	0.2763