Untitled

No description

Report generated at 2019-11-03 18:52:30

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	164853254	151544194
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	163304366	148910512
Mapped(QC-failed)	0	0
% Mapped	99.0600	98.2600
Paired	164853254	151544194
Paired(QC-failed)	0	0
Read1	82426627	75772097
Read1(QC-failed)	0	0
Read2	82426627	75772097
Read2(QC-failed)	0	0
Properly Paired	161397578	146074541
Properly Paired(QC-failed)	0	0
% Properly Paired	97.9000	96.3900
With itself	162546291	147974444
With itself(QC-failed)	0	0
Singletons	758075	936068
Singletons(QC-failed)	0	0
% Singleton	0.4600	0.6200
Diff. Chroms	334006	884346
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	72850395	64801692
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1414558	1185579
Paired Opt. Dupes	16244	14661
% Dupes/100	0.0194	0.0183

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	72849527	64733526
Distinct Read Pairs	71434984	63550092
One Read Pair	70043152	62385083
Two Read Pairs	1369449	1147263
NRF = Distinct/Total	0.9806	0.9817
PBC1 = OnePair/Distinct	0.9805	0.9817
PBC2 = OnePair/TwoPair	51.1470	54.3773

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	142871674	127232226
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	142871674	127232226
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	142871674	127232226
Paired(QC-failed)	0	0
Read1	71435837	63616113
Read1(QC-failed)	0	0
Read2	71435837	63616113
Read2(QC-failed)	0	0
Properly Paired	142871674	127232226
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	142871674	127232226
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	203475
Np	0
N optimal	203475
N conservative	203475
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.	145
Corr. Est. Fragment Len.	0.1721
Phantom Peak	50
Corr. Phantom Peak	0.1735
Argmin. Corr.	1500
Min. Corr.	0.1709
NSC	1.0069
RSC	0.4616

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	0.1374

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	0.2814
AUC	0.4966
CHANCE divergence	0.1065
Elbow Point	0.0000
JS Distance	0.5573
Synthetic AUC	0.5024
Synthetic Elbow Point	0.1452
Synthetic JS Distance	0.2821