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Report generated at 2019-11-03 03:24:49

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	61046626	172537646
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	60209927	169623970
Mapped(QC-failed)	0	0
% Mapped	98.6300	98.3100
Paired	61046626	172537646
Paired(QC-failed)	0	0
Read1	30523313	86268823
Read1(QC-failed)	0	0
Read2	30523313	86268823
Read2(QC-failed)	0	0
Properly Paired	59465814	167189082
Properly Paired(QC-failed)	0	0
% Properly Paired	97.4100	96.9000
With itself	59671568	168209596
With itself(QC-failed)	0	0
Singletons	538359	1414374
Singletons(QC-failed)	0	0
% Singleton	0.8800	0.8200
Diff. Chroms	42282	219888
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	27158241	73240672
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	3640524	4951100
Paired Opt. Dupes	994	3081
% Dupes/100	0.1340	0.0676

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	27156676	73151550
Distinct Read Pairs	23516358	68207516
One Read Pair	20311617	63530412
Two Read Pairs	2816598	4426047
NRF = Distinct/Total	0.8660	0.9324
PBC1 = OnePair/Distinct	0.8637	0.9314
PBC2 = OnePair/TwoPair	7.2114	14.3538

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	47035434	136579144
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	47035434	136579144
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	47035434	136579144
Paired(QC-failed)	0	0
Read1	23517717	68289572
Read1(QC-failed)	0	0
Read2	23517717	68289572
Read2(QC-failed)	0	0
Properly Paired	47035434	136579144
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	47035434	136579144
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	34804
Np	0
N optimal	34804
N conservative	34804
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.	230
Corr. Est. Fragment Len.	0.4603
Phantom Peak	55
Corr. Phantom Peak	0.4058
Argmin. Corr.	1500
Min. Corr.	0.1831
NSC	2.5136
RSC	1.2451

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	0.6409

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	0.0916
AUC	0.4941
CHANCE divergence	0.2326
Elbow Point	0.0000
JS Distance	0.9221
Synthetic AUC	0.4964
Synthetic Elbow Point	0.5953
Synthetic JS Distance	0.6423