Untitled

No description

Report generated at 2021-02-05 11:54:52

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	55823114	99127026
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	55281339	97503736
Mapped(QC-failed)	0	0
% Mapped	99.0300	98.3600
Paired	55823114	99127026
Paired(QC-failed)	0	0
Read1	27911557	49563513
Read1(QC-failed)	0	0
Read2	27911557	49563513
Read2(QC-failed)	0	0
Properly Paired	54767941	95250672
Properly Paired(QC-failed)	0	0
% Properly Paired	98.1100	96.0900
With itself	55000304	96906871
With itself(QC-failed)	0	0
Singletons	281035	596865
Singletons(QC-failed)	0	0
% Singleton	0.5000	0.6000
Diff. Chroms	33183	136587
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	24893208	41226719
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	406951	286989
Paired Opt. Dupes	23583	26725
% Dupes/100	0.0163	0.0070

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	24791577	41202948
Distinct Read Pairs	24397607	40918073
One Read Pair	24031461	40653897
Two Read Pairs	344145	257278
NRF = Distinct/Total	0.9841	0.9931
PBC1 = OnePair/Distinct	0.9850	0.9935
PBC2 = OnePair/TwoPair	69.8295	158.0154

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	48972514	81879460
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	48972514	81879460
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	48972514	81879460
Paired(QC-failed)	0	0
Read1	24486257	40939730
Read1(QC-failed)	0	0
Read2	24486257	40939730
Read2(QC-failed)	0	0
Properly Paired	48972514	81879460
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	48972514	81879460
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	84817
Np	0
N optimal	84817
N conservative	84817
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.	0.2469
Phantom Peak	55
Corr. Phantom Peak	0.2010
Argmin. Corr.	1500
Min. Corr.	0.1660
NSC	1.4869
RSC	2.3115

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	0.4504

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	0.1558
AUC	0.4940
CHANCE divergence	0.1657
Elbow Point	0.0000
JS Distance	0.8149
Synthetic AUC	0.5030
Synthetic Elbow Point	0.3793
Synthetic JS Distance	0.5002