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Report generated at 2021-01-20 09:04:49

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	170547770	193594802
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	167964553	187969057
Mapped(QC-failed)	0	0
% Mapped	98.4900	97.0900
Paired	170547770	193594802
Paired(QC-failed)	0	0
Read1	85273885	96797401
Read1(QC-failed)	0	0
Read2	85273885	96797401
Read2(QC-failed)	0	0
Properly Paired	166295109	184287983
Properly Paired(QC-failed)	0	0
% Properly Paired	97.5100	95.1900
With itself	166912717	185784698
With itself(QC-failed)	0	0
Singletons	1051836	2184359
Singletons(QC-failed)	0	0
% Singleton	0.6200	1.1300
Diff. Chroms	282852	610670
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	73521466	78211821
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1172726	539110
Paired Opt. Dupes	10753	19807
% Dupes/100	0.0160	0.0069

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	73495335	77646644
Distinct Read Pairs	72323194	77172156
One Read Pair	71168286	76701136
Two Read Pairs	1137936	467668
NRF = Distinct/Total	0.9841	0.9939
PBC1 = OnePair/Distinct	0.9840	0.9939
PBC2 = OnePair/TwoPair	62.5416	164.0077

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	144697480	155345422
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	144697480	155345422
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	144697480	155345422
Paired(QC-failed)	0	0
Read1	72348740	77672711
Read1(QC-failed)	0	0
Read2	72348740	77672711
Read2(QC-failed)	0	0
Properly Paired	144697480	155345422
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	144697480	155345422
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	166494
Np	0
N optimal	166494
N conservative	166494
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.	160
Corr. Est. Fragment Len.	0.2105
Phantom Peak	50
Corr. Phantom Peak	0.2094
Argmin. Corr.	1500
Min. Corr.	0.1865
NSC	1.1283
RSC	1.0451

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	0.3889

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	0.2059
AUC	0.4966
CHANCE divergence	0.1017
Elbow Point	0.0000
JS Distance	0.7476
Synthetic AUC	0.5041
Synthetic Elbow Point	0.3212
Synthetic JS Distance	0.4225