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Report generated at 2021-03-31 14:12:13

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	45750040	99621326
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	44153546	94051400
Mapped(QC-failed)	0	0
% Mapped	96.5100	94.4100
Paired	45750040	99621326
Paired(QC-failed)	0	0
Read1	22875020	49810663
Read1(QC-failed)	0	0
Read2	22875020	49810663
Read2(QC-failed)	0	0
Properly Paired	42205814	80789855
Properly Paired(QC-failed)	0	0
% Properly Paired	92.2500	81.1000
With itself	43740610	92478072
With itself(QC-failed)	0	0
Singletons	412936	1573328
Singletons(QC-failed)	0	0
% Singleton	0.9000	1.5800
Diff. Chroms	530013	6482076
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	19075555	36021470
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	840852	466934
Paired Opt. Dupes	963	1238
% Dupes/100	0.0441	0.0130

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	19011920	36005664
Distinct Read Pairs	18183890	35540777
One Read Pair	17391203	35090740
Two Read Pairs	761450	442464
NRF = Distinct/Total	0.9564	0.9871
PBC1 = OnePair/Distinct	0.9564	0.9873
PBC2 = OnePair/TwoPair	22.8396	79.3076

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	36469406	71109072
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	36469406	71109072
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	36469406	71109072
Paired(QC-failed)	0	0
Read1	18234703	35554536
Read1(QC-failed)	0	0
Read2	18234703	35554536
Read2(QC-failed)	0	0
Properly Paired	36469406	71109072
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	36469406	71109072
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	64892
Np	0
N optimal	64892
N conservative	64892
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.	0.1766
Phantom Peak	50
Corr. Phantom Peak	0.1730
Argmin. Corr.	1500
Min. Corr.	0.1621
NSC	1.0891
RSC	1.3278

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	0.1581

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	0.2523
AUC	0.4933
CHANCE divergence	0.1327
Elbow Point	0.0000
JS Distance	0.6151
Synthetic AUC	0.5117
Synthetic Elbow Point	0.1832
Synthetic JS Distance	0.3174