Untitled

No description

Report generated at 2020-05-20 19:34:02

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	126311624	202360194
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	125299851	199280518
Mapped(QC-failed)	0	0
% Mapped	99.2000	98.4800
Paired	126311624	202360194
Paired(QC-failed)	0	0
Read1	63155812	101180097
Read1(QC-failed)	0	0
Read2	63155812	101180097
Read2(QC-failed)	0	0
Properly Paired	123939189	196010642
Properly Paired(QC-failed)	0	0
% Properly Paired	98.1200	96.8600
With itself	124931902	198492948
With itself(QC-failed)	0	0
Singletons	367949	787570
Singletons(QC-failed)	0	0
% Singleton	0.2900	0.3900
Diff. Chroms	54715	212946
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	55701290	87251527
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	2048865	1128860
Paired Opt. Dupes	4200	5289
% Dupes/100	0.0368	0.0129

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	55687631	87198629
Distinct Read Pairs	53639898	86085041
One Read Pair	51665217	85022082
Two Read Pairs	1906253	1031715
NRF = Distinct/Total	0.9632	0.9872
PBC1 = OnePair/Distinct	0.9632	0.9877
PBC2 = OnePair/TwoPair	27.1030	82.4085

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	107304850	172245334
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	107304850	172245334
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	107304850	172245334
Paired(QC-failed)	0	0
Read1	53652425	86122667
Read1(QC-failed)	0	0
Read2	53652425	86122667
Read2(QC-failed)	0	0
Properly Paired	107304850	172245334
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	107304850	172245334
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	172244
Np	0
N optimal	172244
N conservative	172244
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.	0.1785
Phantom Peak	50
Corr. Phantom Peak	0.1771
Argmin. Corr.	1500
Min. Corr.	0.1711
NSC	1.0429
RSC	1.2389

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	0.1549

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	0.2470
AUC	0.4961
CHANCE divergence	0.1338
Elbow Point	0.0000
JS Distance	0.5914
Synthetic AUC	0.5000
Synthetic Elbow Point	0.1320
Synthetic JS Distance	0.3267