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Report generated at 2020-05-13 13:35:54

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	65227664	92901862
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	59481653	87995521
Mapped(QC-failed)	0	0
% Mapped	91.1900	94.7200
Paired	65227664	92901862
Paired(QC-failed)	0	0
Read1	32613832	46450931
Read1(QC-failed)	0	0
Read2	32613832	46450931
Read2(QC-failed)	0	0
Properly Paired	56430606	74516594
Properly Paired(QC-failed)	0	0
% Properly Paired	86.5100	80.2100
With itself	58136216	84730641
With itself(QC-failed)	0	0
Singletons	1345437	3264880
Singletons(QC-failed)	0	0
% Singleton	2.0600	3.5100
Diff. Chroms	782977	6656575
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	23082828	28454434
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	565298	260144
Paired Opt. Dupes	4644	1146
% Dupes/100	0.0245	0.0091

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	23081215	28454249
Distinct Read Pairs	22515943	28194105
One Read Pair	21962184	27936016
Two Read Pairs	542460	256046
NRF = Distinct/Total	0.9755	0.9909
PBC1 = OnePair/Distinct	0.9754	0.9908
PBC2 = OnePair/TwoPair	40.4863	109.1055

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	45035060	56388580
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	45035060	56388580
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	45035060	56388580
Paired(QC-failed)	0	0
Read1	22517530	28194290
Read1(QC-failed)	0	0
Read2	22517530	28194290
Read2(QC-failed)	0	0
Properly Paired	45035060	56388580
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	45035060	56388580
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	172008
Np	0
N optimal	172008
N conservative	172008
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.	270
Corr. Est. Fragment Len.	0.2357
Phantom Peak	50
Corr. Phantom Peak	0.2403
Argmin. Corr.	1500
Min. Corr.	0.1869
NSC	1.2607
RSC	0.9128

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	0.3828

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	0.1090
AUC	0.4939
CHANCE divergence	0.4716
Elbow Point	0.0000
JS Distance	0.7276
Synthetic AUC	0.5014
Synthetic Elbow Point	0.1391
Synthetic JS Distance	0.4444