Untitled

No description

Report generated at 2019-10-13 05:00:47

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	90871374	95805330
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	85835879	91174239
Mapped(QC-failed)	0	0
% Mapped	94.4600	95.1700
Paired	90871374	95805330
Paired(QC-failed)	0	0
Read1	45435687	47902665
Read1(QC-failed)	0	0
Read2	45435687	47902665
Read2(QC-failed)	0	0
Properly Paired	82075785	87029614
Properly Paired(QC-failed)	0	0
% Properly Paired	90.3200	90.8400
With itself	82793963	88132031
With itself(QC-failed)	0	0
Singletons	3041916	3042208
Singletons(QC-failed)	0	0
% Singleton	3.3500	3.1800
Diff. Chroms	313872	677774
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	33631279	36289675
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	4586771	346498
Paired Opt. Dupes	4403	3852
% Dupes/100	0.1364	0.0095

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	33630311	36261516
Distinct Read Pairs	29043664	35915541
One Read Pair	25082534	35581631
Two Read Pairs	3419987	326616
NRF = Distinct/Total	0.8636	0.9905
PBC1 = OnePair/Distinct	0.8636	0.9907
PBC2 = OnePair/TwoPair	7.3341	108.9403

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	58089016	71886354
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	58089016	71886354
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	58089016	71886354
Paired(QC-failed)	0	0
Read1	29044508	35943177
Read1(QC-failed)	0	0
Read2	29044508	35943177
Read2(QC-failed)	0	0
Properly Paired	58089016	71886354
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	58089016	71886354
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	16092
Np	0
N optimal	16092
N conservative	16092
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.	125
Corr. Est. Fragment Len.	0.1717
Phantom Peak	50
Corr. Phantom Peak	0.1892
Argmin. Corr.	1500
Min. Corr.	0.1665
NSC	1.0313
RSC	0.2299

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	0.0079

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	0.3130
AUC	0.4946
CHANCE divergence	0.1055
Elbow Point	0.0000
JS Distance	0.5223
Synthetic AUC	0.5040
Synthetic Elbow Point	0.0252
Synthetic JS Distance	0.2216