Untitled

No description

Report generated at 2020-05-27 04:24:12

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	104968594	199291734
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	55383363	192758941
Mapped(QC-failed)	0	0
% Mapped	52.7600	96.7200
Paired	104968594	199291734
Paired(QC-failed)	0	0
Read1	52484297	99645867
Read1(QC-failed)	0	0
Read2	52484297	99645867
Read2(QC-failed)	0	0
Properly Paired	53911895	190351557
Properly Paired(QC-failed)	0	0
% Properly Paired	51.3600	95.5100
With itself	54235195	191591556
With itself(QC-failed)	0	0
Singletons	1148168	1167385
Singletons(QC-failed)	0	0
% Singleton	1.0900	0.5900
Diff. Chroms	77206	372346
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	23485564	82947353
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	330462	947217
Paired Opt. Dupes	1312	3959
% Dupes/100	0.0141	0.0114

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	23480307	82921397
Distinct Read Pairs	23149925	81974628
One Read Pair	22824209	81039698
Two Read Pairs	321132	923351
NRF = Distinct/Total	0.9859	0.9886
PBC1 = OnePair/Distinct	0.9859	0.9886
PBC2 = OnePair/TwoPair	71.0742	87.7669

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	46310204	164000272
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	46310204	164000272
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	46310204	164000272
Paired(QC-failed)	0	0
Read1	23155102	82000136
Read1(QC-failed)	0	0
Read2	23155102	82000136
Read2(QC-failed)	0	0
Properly Paired	46310204	164000272
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	46310204	164000272
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	32589
Np	0
N optimal	32589
N conservative	32589
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.	140
Corr. Est. Fragment Len.	0.1849
Phantom Peak	50
Corr. Phantom Peak	0.2320
Argmin. Corr.	1500
Min. Corr.	0.1632
NSC	1.1329
RSC	0.3153

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	0.1189

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	0.2427
AUC	0.4940
CHANCE divergence	0.1427
Elbow Point	0.0000
JS Distance	0.6151
Synthetic AUC	0.5027
Synthetic Elbow Point	0.1559
Synthetic JS Distance	0.3315