Untitled

No description

Report generated at 2020-05-13 13:13:14

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	68077970	78498540
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	54008904	74844187
Mapped(QC-failed)	0	0
% Mapped	79.3300	95.3400
Paired	68077970	78498540
Paired(QC-failed)	0	0
Read1	34038985	39249270
Read1(QC-failed)	0	0
Read2	34038985	39249270
Read2(QC-failed)	0	0
Properly Paired	50683928	67679938
Properly Paired(QC-failed)	0	0
% Properly Paired	74.4500	86.2200
With itself	51670226	72834832
With itself(QC-failed)	0	0
Singletons	2338678	2009355
Singletons(QC-failed)	0	0
% Singleton	3.4400	2.5600
Diff. Chroms	658302	3424690
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	22466317	28005715
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	996415	528129
Paired Opt. Dupes	2071	2652
% Dupes/100	0.0444	0.0189

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	22458061	28000939
Distinct Read Pairs	21462002	27472963
One Read Pair	20503442	26953277
Two Read Pairs	922333	511504
NRF = Distinct/Total	0.9556	0.9811
PBC1 = OnePair/Distinct	0.9553	0.9811
PBC2 = OnePair/TwoPair	22.2300	52.6942

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	42939804	54955172
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	42939804	54955172
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	42939804	54955172
Paired(QC-failed)	0	0
Read1	21469902	27477586
Read1(QC-failed)	0	0
Read2	21469902	27477586
Read2(QC-failed)	0	0
Properly Paired	42939804	54955172
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	42939804	54955172
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	28125
Np	0
N optimal	28125
N conservative	28125
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.	260
Corr. Est. Fragment Len.	0.3538
Phantom Peak	50
Corr. Phantom Peak	0.3095
Argmin. Corr.	1500
Min. Corr.	0.1799
NSC	1.9671
RSC	1.3419

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	0.0870
AUC	0.4937
CHANCE divergence	0.4568
Elbow Point	0.0000
JS Distance	0.7704
Synthetic AUC	0.5109
Synthetic Elbow Point	0.3841
Synthetic JS Distance	0.5434