Untitled

No description

Report generated at 2020-07-10 02:22:19

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	154036472	131098510
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	152766955	129303333
Mapped(QC-failed)	0	0
% Mapped	99.1800	98.6300
Paired	154036472	131098510
Paired(QC-failed)	0	0
Read1	77018236	65549255
Read1(QC-failed)	0	0
Read2	77018236	65549255
Read2(QC-failed)	0	0
Properly Paired	151590183	124505591
Properly Paired(QC-failed)	0	0
% Properly Paired	98.4100	94.9700
With itself	152125760	128591289
With itself(QC-failed)	0	0
Singletons	641195	712044
Singletons(QC-failed)	0	0
% Singleton	0.4200	0.5400
Diff. Chroms	71580	201152
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	70388421	54582151
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1162388	411333
Paired Opt. Dupes	4169	2925
% Dupes/100	0.0165	0.0075

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	70370894	54552078
Distinct Read Pairs	69209932	54144593
One Read Pair	68082361	53754757
Two Read Pairs	1097739	382787
NRF = Distinct/Total	0.9835	0.9925
PBC1 = OnePair/Distinct	0.9837	0.9928
PBC2 = OnePair/TwoPair	62.0205	140.4299

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	138452066	108341636
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	138452066	108341636
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	138452066	108341636
Paired(QC-failed)	0	0
Read1	69226033	54170818
Read1(QC-failed)	0	0
Read2	69226033	54170818
Read2(QC-failed)	0	0
Properly Paired	138452066	108341636
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	138452066	108341636
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	225778
Np	0
N optimal	225778
N conservative	225778
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.	0.1827
Phantom Peak	45
Corr. Phantom Peak	0.1792
Argmin. Corr.	1500
Min. Corr.	0.1711
NSC	1.0680
RSC	1.4365

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	0.4532

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	0.2063
AUC	0.4966
CHANCE divergence	0.1043
Elbow Point	0.0000
JS Distance	0.7605
Synthetic AUC	0.5049
Synthetic Elbow Point	0.2851
Synthetic JS Distance	0.4086