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Report generated at 2020-05-03 09:41:31

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	81113800	136790320
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	80203509	132380851
Mapped(QC-failed)	0	0
% Mapped	98.8800	96.7800
Paired	81113800	136790320
Paired(QC-failed)	0	0
Read1	40556900	68395160
Read1(QC-failed)	0	0
Read2	40556900	68395160
Read2(QC-failed)	0	0
Properly Paired	79555478	130092484
Properly Paired(QC-failed)	0	0
% Properly Paired	98.0800	95.1000
With itself	80012708	131677058
With itself(QC-failed)	0	0
Singletons	190801	703793
Singletons(QC-failed)	0	0
% Singleton	0.2400	0.5100
Diff. Chroms	254281	604160
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	37020786	56350303
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	200560	241110
Paired Opt. Dupes	1862	2032
% Dupes/100	0.0054	0.0043

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	37019939	56321306
Distinct Read Pairs	36819381	56080639
One Read Pair	36619659	55842177
Two Read Pairs	198893	236573
NRF = Distinct/Total	0.9946	0.9957
PBC1 = OnePair/Distinct	0.9946	0.9957
PBC2 = OnePair/TwoPair	184.1174	236.0463

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	73640452	112218386
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	73640452	112218386
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	73640452	112218386
Paired(QC-failed)	0	0
Read1	36820226	56109193
Read1(QC-failed)	0	0
Read2	36820226	56109193
Read2(QC-failed)	0	0
Properly Paired	73640452	112218386
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	73640452	112218386
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	176577
Np	0
N optimal	176577
N conservative	176577
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.	135
Corr. Est. Fragment Len.	0.2239
Phantom Peak	50
Corr. Phantom Peak	0.2171
Argmin. Corr.	1500
Min. Corr.	0.1892
NSC	1.1832
RSC	1.2410

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	0.4310

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	0.1381
AUC	0.4953
CHANCE divergence	0.2312
Elbow Point	0.0000
JS Distance	0.7563
Synthetic AUC	0.5042
Synthetic Elbow Point	0.2868
Synthetic JS Distance	0.4924