Untitled

No description

Report generated at 2020-05-14 01:49:46

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	43273052	75348014
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	41659024	72639776
Mapped(QC-failed)	0	0
% Mapped	96.2700	96.4100
Paired	43273052	75348014
Paired(QC-failed)	0	0
Read1	21636526	37674007
Read1(QC-failed)	0	0
Read2	21636526	37674007
Read2(QC-failed)	0	0
Properly Paired	41122232	70114889
Properly Paired(QC-failed)	0	0
% Properly Paired	95.0300	93.0500
With itself	41495310	71857749
With itself(QC-failed)	0	0
Singletons	163714	782027
Singletons(QC-failed)	0	0
% Singleton	0.3800	1.0400
Diff. Chroms	221437	1077494
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	18805219	30082479
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	61884	117823
Paired Opt. Dupes	385	447
% Dupes/100	0.0033	0.0039

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	18803854	30056490
Distinct Read Pairs	18741977	29938925
One Read Pair	18680260	29821774
Two Read Pairs	61557	116742
NRF = Distinct/Total	0.9967	0.9961
PBC1 = OnePair/Distinct	0.9967	0.9961
PBC2 = OnePair/TwoPair	303.4628	255.4503

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	37486670	59929312
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	37486670	59929312
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	37486670	59929312
Paired(QC-failed)	0	0
Read1	18743335	29964656
Read1(QC-failed)	0	0
Read2	18743335	29964656
Read2(QC-failed)	0	0
Properly Paired	37486670	59929312
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	37486670	59929312
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	94629
Np	0
N optimal	94629
N conservative	94629
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.	175
Corr. Est. Fragment Len.	0.1994
Phantom Peak	50
Corr. Phantom Peak	0.2010
Argmin. Corr.	1500
Min. Corr.	0.1841
NSC	1.0833
RSC	0.9082

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	0.2801

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	0.1766
AUC	0.4934
CHANCE divergence	0.2054
Elbow Point	0.0000
JS Distance	0.7217
Synthetic AUC	0.5040
Synthetic Elbow Point	0.1800
Synthetic JS Distance	0.4174