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Report generated at 2020-07-10 05:14:08

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	160095336	136244046
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	158333946	134324819
Mapped(QC-failed)	0	0
% Mapped	98.9000	98.5900
Paired	160095336	136244046
Paired(QC-failed)	0	0
Read1	80047668	68122023
Read1(QC-failed)	0	0
Read2	80047668	68122023
Read2(QC-failed)	0	0
Properly Paired	156711061	131038518
Properly Paired(QC-failed)	0	0
% Properly Paired	97.8900	96.1800
With itself	157583336	133591989
With itself(QC-failed)	0	0
Singletons	750610	732830
Singletons(QC-failed)	0	0
% Singleton	0.4700	0.5400
Diff. Chroms	84767	172253
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	71690512	57476635
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1050901	368798
Paired Opt. Dupes	3672	2996
% Dupes/100	0.0147	0.0064

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	71676346	57449552
Distinct Read Pairs	70626249	57083273
One Read Pair	69598116	56730634
Two Read Pairs	1009445	346986
NRF = Distinct/Total	0.9853	0.9936
PBC1 = OnePair/Distinct	0.9854	0.9938
PBC2 = OnePair/TwoPair	68.9469	163.4955

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	141279222	114215674
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	141279222	114215674
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	141279222	114215674
Paired(QC-failed)	0	0
Read1	70639611	57107837
Read1(QC-failed)	0	0
Read2	70639611	57107837
Read2(QC-failed)	0	0
Properly Paired	141279222	114215674
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	141279222	114215674
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	223034
Np	0
N optimal	223034
N conservative	223034
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.	0.1753
Phantom Peak	50
Corr. Phantom Peak	0.1756
Argmin. Corr.	1500
Min. Corr.	0.1687
NSC	1.0393
RSC	0.9659

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	0.2565

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	0.2734
AUC	0.4966
CHANCE divergence	0.0996
Elbow Point	0.0000
JS Distance	0.6697
Synthetic AUC	0.5030
Synthetic Elbow Point	0.1666
Synthetic JS Distance	0.2944