Untitled

No description

Report generated at 2022-09-02 23:21:19

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	180895024	50889240
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	165249596	50058651
Mapped(QC-failed)	0	0
% Mapped	91.3500	98.3700
Paired	180895024	50889240
Paired(QC-failed)	0	0
Read1	90447512	25444620
Read1(QC-failed)	0	0
Read2	90447512	25444620
Read2(QC-failed)	0	0
Properly Paired	163673999	49717706
Properly Paired(QC-failed)	0	0
% Properly Paired	90.4800	97.7000
With itself	164519914	49906072
With itself(QC-failed)	0	0
Singletons	729682	152579
Singletons(QC-failed)	0	0
% Singleton	0.4000	0.3000
Diff. Chroms	196262	67298
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	73113113	22060886
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1712455	95014
Paired Opt. Dupes	6622	6064
% Dupes/100	0.0234	0.0043

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	73109408	22024154
Distinct Read Pairs	71397057	21929940
One Read Pair	69723723	21836362
Two Read Pairs	1636293	93000
NRF = Distinct/Total	0.9766	0.9957
PBC1 = OnePair/Distinct	0.9766	0.9957
PBC2 = OnePair/TwoPair	42.6108	234.7996

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	142801316	43931744
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	142801316	43931744
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	142801316	43931744
Paired(QC-failed)	0	0
Read1	71400658	21965872
Read1(QC-failed)	0	0
Read2	71400658	21965872
Read2(QC-failed)	0	0
Properly Paired	142801316	43931744
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	142801316	43931744
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	100057
Np	0
N optimal	100057
N conservative	100057
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.	135
Corr. Est. Fragment Len.	0.1806
Phantom Peak	50
Corr. Phantom Peak	0.1936
Argmin. Corr.	1500
Min. Corr.	0.1752
NSC	1.0306
RSC	0.2907

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	0.0788

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	0.2809
AUC	0.4966
CHANCE divergence	0.1025
Elbow Point	0.0000
JS Distance	0.6425
Synthetic AUC	0.4996
Synthetic Elbow Point	0.0822
Synthetic JS Distance	0.2811