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Report generated at 2022-09-02 20:52:01

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	160327598	50889240
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	152933851	50058651
Mapped(QC-failed)	0	0
% Mapped	95.3900	98.3700
Paired	160327598	50889240
Paired(QC-failed)	0	0
Read1	80163799	25444620
Read1(QC-failed)	0	0
Read2	80163799	25444620
Read2(QC-failed)	0	0
Properly Paired	151640305	49717706
Properly Paired(QC-failed)	0	0
% Properly Paired	94.5800	97.7000
With itself	152393761	49906072
With itself(QC-failed)	0	0
Singletons	540090	152579
Singletons(QC-failed)	0	0
% Singleton	0.3400	0.3000
Diff. Chroms	121750	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	67618863	22060886
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	1465874	95014
Paired Opt. Dupes	7047	6064
% Dupes/100	0.0217	0.0043

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	67614857	22024154
Distinct Read Pairs	66149081	21929940
One Read Pair	64712755	21836362
Two Read Pairs	1407818	93000
NRF = Distinct/Total	0.9783	0.9957
PBC1 = OnePair/Distinct	0.9783	0.9957
PBC2 = OnePair/TwoPair	45.9667	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	132305978	43931744
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	132305978	43931744
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	132305978	43931744
Paired(QC-failed)	0	0
Read1	66152989	21965872
Read1(QC-failed)	0	0
Read2	66152989	21965872
Read2(QC-failed)	0	0
Properly Paired	132305978	43931744
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	132305978	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	38892
Np	0
N optimal	38892
N conservative	38892
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.	130
Corr. Est. Fragment Len.	0.1779
Phantom Peak	50
Corr. Phantom Peak	0.1890
Argmin. Corr.	1500
Min. Corr.	0.1729
NSC	1.0288
RSC	0.3089

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	0.0217

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	0.3121
AUC	0.4965
CHANCE divergence	0.1018
Elbow Point	0.0000
JS Distance	0.5472
Synthetic AUC	0.4986
Synthetic Elbow Point	0.0338
Synthetic JS Distance	0.2288