Untitled

No description

Report generated at 2020-07-14 23:25:28

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	69774438	161541774
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	65623680	157457809
Mapped(QC-failed)	0	0
% Mapped	94.0500	97.4700
Paired	69774438	161541774
Paired(QC-failed)	0	0
Read1	34887219	80770887
Read1(QC-failed)	0	0
Read2	34887219	80770887
Read2(QC-failed)	0	0
Properly Paired	64746457	154923170
Properly Paired(QC-failed)	0	0
% Properly Paired	92.7900	95.9000
With itself	65287028	156706856
With itself(QC-failed)	0	0
Singletons	336652	750953
Singletons(QC-failed)	0	0
% Singleton	0.4800	0.4600
Diff. Chroms	333987	1033935
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	29370468	67677947
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	92278	255439
Paired Opt. Dupes	672	1182
% Dupes/100	0.0031	0.0038

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	29368697	67623047
Distinct Read Pairs	29276425	67368479
One Read Pair	29184343	67115243
Two Read Pairs	91893	251987
NRF = Distinct/Total	0.9969	0.9962
PBC1 = OnePair/Distinct	0.9969	0.9962
PBC2 = OnePair/TwoPair	317.5905	266.3441

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	58556380	134845016
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	58556380	134845016
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	58556380	134845016
Paired(QC-failed)	0	0
Read1	29278190	67422508
Read1(QC-failed)	0	0
Read2	29278190	67422508
Read2(QC-failed)	0	0
Properly Paired	58556380	134845016
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	58556380	134845016
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	128891
Np	0
N optimal	128891
N conservative	128891
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.	140
Corr. Est. Fragment Len.	0.1867
Phantom Peak	50
Corr. Phantom Peak	0.1969
Argmin. Corr.	1500
Min. Corr.	0.1768
NSC	1.0559
RSC	0.4921

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	0.2252

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	0.2301
AUC	0.4947
CHANCE divergence	0.1268
Elbow Point	0.0000
JS Distance	0.6655
Synthetic AUC	0.5084
Synthetic Elbow Point	0.2125
Synthetic JS Distance	0.3544