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Report generated at 2020-05-20 08:11:08

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	72806404	118522358
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	71489245	116921431
Mapped(QC-failed)	0	0
% Mapped	98.1900	98.6500
Paired	72806404	118522358
Paired(QC-failed)	0	0
Read1	36403202	59261179
Read1(QC-failed)	0	0
Read2	36403202	59261179
Read2(QC-failed)	0	0
Properly Paired	69553942	111554301
Properly Paired(QC-failed)	0	0
% Properly Paired	95.5300	94.1200
With itself	71211043	116290293
With itself(QC-failed)	0	0
Singletons	278202	631138
Singletons(QC-failed)	0	0
% Singleton	0.3800	0.5300
Diff. Chroms	57365	155795
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	30892019	49637685
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	2455317	399141
Paired Opt. Dupes	2221	4222
% Dupes/100	0.0795	0.0080

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	30887520	49626135
Distinct Read Pairs	28432550	49227867
One Read Pair	26159961	48851116
Two Read Pairs	2121456	369226
NRF = Distinct/Total	0.9205	0.9920
PBC1 = OnePair/Distinct	0.9201	0.9923
PBC2 = OnePair/TwoPair	12.3311	132.3068

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	56873404	98477088
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	56873404	98477088
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	56873404	98477088
Paired(QC-failed)	0	0
Read1	28436702	49238544
Read1(QC-failed)	0	0
Read2	28436702	49238544
Read2(QC-failed)	0	0
Properly Paired	56873404	98477088
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	56873404	98477088
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	59258
Np	0
N optimal	59258
N conservative	59258
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.	150
Corr. Est. Fragment Len.	0.1814
Phantom Peak	50
Corr. Phantom Peak	0.1762
Argmin. Corr.	1500
Min. Corr.	0.1628
NSC	1.1139
RSC	1.3843

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	0.0757

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	0.2811
AUC	0.4946
CHANCE divergence	0.1170
Elbow Point	0.0000
JS Distance	0.5636
Synthetic AUC	0.4966
Synthetic Elbow Point	0.1223
Synthetic JS Distance	0.2753