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Report generated at 2020-05-04 15:28:03

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	187649404	185333438
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	142387372	179322251
Mapped(QC-failed)	0	0
% Mapped	75.8800	96.7600
Paired	187649404	185333438
Paired(QC-failed)	0	0
Read1	93824702	92666719
Read1(QC-failed)	0	0
Read2	93824702	92666719
Read2(QC-failed)	0	0
Properly Paired	139057224	177236646
Properly Paired(QC-failed)	0	0
% Properly Paired	74.1000	95.6300
With itself	140222671	178226497
With itself(QC-failed)	0	0
Singletons	2164701	1095754
Singletons(QC-failed)	0	0
% Singleton	1.1500	0.5900
Diff. Chroms	284338	321793
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	55889542	76478349
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	572975	700148
Paired Opt. Dupes	2952	1938
% Dupes/100	0.0103	0.0092

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	55886880	76473537
Distinct Read Pairs	55313928	75773440
One Read Pair	54750662	75080087
Two Read Pairs	554422	686711
NRF = Distinct/Total	0.9897	0.9908
PBC1 = OnePair/Distinct	0.9898	0.9909
PBC2 = OnePair/TwoPair	98.7527	109.3329

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	110633134	151556402
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	110633134	151556402
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	110633134	151556402
Paired(QC-failed)	0	0
Read1	55316567	75778201
Read1(QC-failed)	0	0
Read2	55316567	75778201
Read2(QC-failed)	0	0
Properly Paired	110633134	151556402
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	110633134	151556402
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	180726
Np	0
N optimal	180726
N conservative	180726
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.	110
Corr. Est. Fragment Len.	0.1962
Phantom Peak	50
Corr. Phantom Peak	0.2398
Argmin. Corr.	1500
Min. Corr.	0.1840
NSC	1.0662
RSC	0.2183

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	0.1391

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	0.2573
AUC	0.4961
CHANCE divergence	0.1011
Elbow Point	0.0000
JS Distance	0.6163
Synthetic AUC	0.5017
Synthetic Elbow Point	0.0968
Synthetic JS Distance	0.3206