Untitled

No description

Report generated at 2019-10-13 04:25:31

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	94843886	87738496
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	93327529	86229239
Mapped(QC-failed)	0	0
% Mapped	98.4000	98.2800
Paired	94843886	87738496
Paired(QC-failed)	0	0
Read1	47421943	43869248
Read1(QC-failed)	0	0
Read2	47421943	43869248
Read2(QC-failed)	0	0
Properly Paired	92203586	85110249
Properly Paired(QC-failed)	0	0
% Properly Paired	97.2200	97.0000
With itself	92738192	85780325
With itself(QC-failed)	0	0
Singletons	589337	448914
Singletons(QC-failed)	0	0
% Singleton	0.6200	0.5100
Diff. Chroms	147258	132532
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	40733030	37459366
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	2591254	810815
Paired Opt. Dupes	14887	14619
% Dupes/100	0.0636	0.0216

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	40731923	37393436
Distinct Read Pairs	38140749	36585669
One Read Pair	35701741	35798621
Two Read Pairs	2295441	769016
NRF = Distinct/Total	0.9364	0.9784
PBC1 = OnePair/Distinct	0.9361	0.9785
PBC2 = OnePair/TwoPair	15.5533	46.5512

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	76283552	73297102
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	76283552	73297102
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	76283552	73297102
Paired(QC-failed)	0	0
Read1	38141776	36648551
Read1(QC-failed)	0	0
Read2	38141776	36648551
Read2(QC-failed)	0	0
Properly Paired	76283552	73297102
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	76283552	73297102
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	218925
Np	0
N optimal	218925
N conservative	218925
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.	0.1747
Phantom Peak	50
Corr. Phantom Peak	0.1811
Argmin. Corr.	1500
Min. Corr.	0.1714
NSC	1.0192
RSC	0.3399

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	0.2299

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	0.2455
AUC	0.4953
CHANCE divergence	0.1071
Elbow Point	0.0000
JS Distance	0.6794
Synthetic AUC	0.5074
Synthetic Elbow Point	0.1628
Synthetic JS Distance	0.3328