Untitled

No description

Report generated at 2020-05-02 12:31:15

Pipeline type: Histone ChIP-Seq

Peak caller: MACS2

Alignment

Flagstat (raw BAM)

	rep1	ctl1
Total	34248740	49040330
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	24000806	43899885
Mapped(QC-failed)	0	0
% Mapped	70.0800	89.5200
Paired	34248740	49040330
Paired(QC-failed)	0	0
Read1	17124370	24520165
Read1(QC-failed)	0	0
Read2	17124370	24520165
Read2(QC-failed)	0	0
Properly Paired	18932785	30373686
Properly Paired(QC-failed)	0	0
% Properly Paired	55.2800	61.9400
With itself	22520466	41523513
With itself(QC-failed)	0	0
Singletons	1480340	2376372
Singletons(QC-failed)	0	0
% Singleton	4.3200	4.8500
Diff. Chroms	2627093	7654539
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	7347903	10159811
Unmapped Reads	0	0
Unpaired Dupes	0	0
Paired Dupes	220480	264224
Paired Opt. Dupes	557	1355
% Dupes/100	0.0300	0.0260

Library complexity (filtered non-mito BAM)

	rep1	ctl1
Total Read Pairs	7346872	10158782
Distinct Read Pairs	7126418	9894586
One Read Pair	6911490	9636269
Two Read Pairs	209528	252589
NRF = Distinct/Total	0.9700	0.9740
PBC1 = OnePair/Distinct	0.9698	0.9739
PBC2 = OnePair/TwoPair	32.9860	38.1500

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	14254846	19791174
Total(QC-failed)	0	0
Dupes	0	0
Dupes(QC-failed)	0	0
Mapped	14254846	19791174
Mapped(QC-failed)	0	0
% Mapped	100.0000	100.0000
Paired	14254846	19791174
Paired(QC-failed)	0	0
Read1	7127423	9895587
Read1(QC-failed)	0	0
Read2	7127423	9895587
Read2(QC-failed)	0	0
Properly Paired	14254846	19791174
Properly Paired(QC-failed)	0	0
% Properly Paired	100.0000	100.0000
With itself	14254846	19791174
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	29574
Np	0
N optimal	29574
N conservative	29574
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 (11M)

	rep1
Reads	11412467
Est. Fragment Len.	225
Corr. Est. Fragment Len.	0.1638
Phantom Peak	50
Corr. Phantom Peak	0.1783
Argmin. Corr.	1500
Min. Corr.	0.1444
NSC	1.1349
RSC	0.5739

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	0.1011

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	0.1331
AUC	0.4891
CHANCE divergence	0.4540
Elbow Point	0.0000
JS Distance	0.7183
Synthetic AUC	0.5012
Synthetic Elbow Point	0.1177
Synthetic JS Distance	0.3694