Untitled

No description

Report generated at 2019-10-30 20:50:32

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	78578740	186328420
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	68862333	182595069
Mapped(QC-failed)	0	0
% Mapped	87.6300	98.0000
Paired	78578740	186328420
Paired(QC-failed)	0	0
Read1	39289370	93164210
Read1(QC-failed)	0	0
Read2	39289370	93164210
Read2(QC-failed)	0	0
Properly Paired	67606159	174653171
Properly Paired(QC-failed)	0	0
% Properly Paired	86.0400	93.7300
With itself	68100410	180890277
With itself(QC-failed)	0	0
Singletons	761923	1704792
Singletons(QC-failed)	0	0
% Singleton	0.9700	0.9100
Diff. Chroms	208749	4529429
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	29087361	73010674
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	11316369	6709019
Paired Opt. Dupes	5650	13379
% Dupes/100	0.3890	0.0919

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	29083476	72957325
Distinct Read Pairs	17768571	66253408
One Read Pair	10341048	60053338
Two Read Pairs	4747871	5734419
NRF = Distinct/Total	0.6110	0.9081
PBC1 = OnePair/Distinct	0.5820	0.9064
PBC2 = OnePair/TwoPair	2.1780	10.4724

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	35541984	132603310
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	35541984	132603310
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	35541984	132603310
Paired(QC-failed)	0	0
Read1	17770992	66301655
Read1(QC-failed)	0	0
Read2	17770992	66301655
Read2(QC-failed)	0	0
Properly Paired	35541984	132603310
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	35541984	132603310
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	79428
Np	0
N optimal	79428
N conservative	79428
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.	170
Corr. Est. Fragment Len.	0.1576
Phantom Peak	50
Corr. Phantom Peak	0.1579
Argmin. Corr.	1500
Min. Corr.	0.1430
NSC	1.1020
RSC	0.9817

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	0.2479

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	0.2180
AUC	0.4917
CHANCE divergence	0.1583
Elbow Point	0.0000
JS Distance	0.6740
Synthetic AUC	0.4977
Synthetic Elbow Point	0.2759
Synthetic JS Distance	0.3634