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Report generated at 2020-05-13 02:57:26

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	58646462	44822058
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	55884833	42270277
Mapped(QC-failed)	0	0
% Mapped	95.2900	94.3100
Paired	58646462	44822058
Paired(QC-failed)	0	0
Read1	29323231	22411029
Read1(QC-failed)	0	0
Read2	29323231	22411029
Read2(QC-failed)	0	0
Properly Paired	53929361	39106937
Properly Paired(QC-failed)	0	0
% Properly Paired	91.9600	87.2500
With itself	55097242	41498214
With itself(QC-failed)	0	0
Singletons	787591	772063
Singletons(QC-failed)	0	0
% Singleton	1.3400	1.7200
Diff. Chroms	844962	1698756
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	24044852	17000414
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	224390	184749
Paired Opt. Dupes	2159	1571
% Dupes/100	0.0093	0.0109

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	24044552	16997550
Distinct Read Pairs	23820164	16812850
One Read Pair	23597610	16629884
Two Read Pairs	220735	181248
NRF = Distinct/Total	0.9907	0.9891
PBC1 = OnePair/Distinct	0.9907	0.9891
PBC2 = OnePair/TwoPair	106.9047	91.7521

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	47640924	33631330
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	47640924	33631330
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	47640924	33631330
Paired(QC-failed)	0	0
Read1	23820462	16815665
Read1(QC-failed)	0	0
Read2	23820462	16815665
Read2(QC-failed)	0	0
Properly Paired	47640924	33631330
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	47640924	33631330
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	124331
Np	0
N optimal	124331
N conservative	124331
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.	275
Corr. Est. Fragment Len.	0.2117
Phantom Peak	50
Corr. Phantom Peak	0.2016
Argmin. Corr.	1500
Min. Corr.	0.1921
NSC	1.1020
RSC	2.0692

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	0.3450

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	0.1224
AUC	0.4941
CHANCE divergence	0.4086
Elbow Point	0.0000
JS Distance	0.7198
Synthetic AUC	0.4971
Synthetic Elbow Point	0.1852
Synthetic JS Distance	0.4413