Open chromatin assays are known to have significant GC bias. Please take this
into consideration as necessary.
Library complexity quality metrics
Library complexity (filtered non-mito BAM)
rep1
rep2
rep3
ctl1
Total Fragments
3351613
3932005
1340189
6809810
Distinct Fragments
2694229
3163685
1179037
6459250
Positions with Two Read
435690
504900
124688
301612
NRF = Distinct/Total
0.80386
0.804598
0.879754
0.948521
PBC1 = OneRead/Distinct
0.802285
0.804211
0.880011
0.950279
PBC2 = OneRead/TwoRead
4.961188
5.039156
8.321298
20.350954
Mitochondrial reads are filtered out by default.
The non-redundant fraction (NRF) is the fraction of non-redundant mapped reads
in a dataset; it is the ratio between the number of positions in the genome
that uniquely mapped reads map to and the total number of uniquely mappable
reads. The NRF should be > 0.8. The PBC1 is the ratio of genomic locations
with EXACTLY one read pair over the genomic locations with AT LEAST one read
pair. PBC1 is the primary measure, and the PBC1 should be close to 1.
Provisionally 0-0.5 is severe bottlenecking, 0.5-0.8 is moderate bottlenecking,
0.8-0.9 is mild bottlenecking, and 0.9-1.0 is no bottlenecking. The PBC2 is
the ratio of genomic locations with EXACTLY one read pair over the genomic
locations with EXACTLY two read pairs. The PBC2 should be significantly
greater than 1. See more details at
the ENCODE portal standard for ChIP-Seq pipeline
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
Replication quality metrics
IDR (Irreproducible Discovery Rate) plots
Reproducibility QC and peak detection statistics
overlap
idr
Nt
25749
5606
N1
18358
3539
N2
20155
3501
N3
10619
1933
Np
27533
6510
N optimal
27533
6510
N conservative
25749
5606
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep2
rep1_vs_rep2
Rescue Ratio
1.0692842440483126
1.1612557973599715
Self Consistency Ratio
1.898012995573971
1.8308329022245216
Reproducibility Test
pass
pass
Reproducibility QC
N1: Replicate 1 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
N2: Replicate 2 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
Ni: Replicate i self-consistent peaks (comparing two pseudoreplicates generated by subsampling RepX reads)
Nt: True Replicate consistent peaks (comparing true replicates Rep1 vs Rep2)
Np: Pooled-pseudoreplicate consistent 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'
Number of raw peaks
rep1
rep2
rep3
Number of peaks
39560
42958
29599
Top 300000 raw peaks from spp with FDR 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
idr_opt
overlap_opt
Min size
115.0
119.0
111.0
115.0
115.0
25 percentile
460.0
476.0
444.0
353.0
460.0
50 percentile (median)
460.0
476.0
444.0
460.0
460.0
75 percentile
460.0
476.0
444.0
460.0
460.0
Max size
1298.0
1347.0
765.0
13297.0
13297.0
Mean
454.67692113245704
471.70478141440475
440.28281360856784
457.83533026113673
459.07779755202847
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
3351613
3932005
1340189
Estimated Fragment Length
175
175
175
Cross-correlation at Estimated Fragment Length
0.527197078144913
0.556116560556695
0.355765269410071
Phantom Peak
40
40
40
Cross-correlation at Phantom Peak
0.5033604
0.5377483
0.334238
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.4615721
0.5021741
0.2981634
NSC (Normalized Strand Cross-correlation coeff.)
1.142177
1.107418
1.193189
RSC (Relative Strand Cross-correlation coeff.)
1.570415
1.516336
1.596744
Performed on subsampled (15000000) reads mapped from FASTQs that are trimmed to 50.
Such FASTQ trimming and subsampling reads are for cross-corrleation analysis only.
Untrimmed FASTQs are used for all the other analyses.
NOTE1: For SE datasets, reads from replicates are randomly subsampled to 15000000.
NOTE2: For PE datasets, the first end (R1) of each read-pair is selected and trimmed to 50 the reads are then randomly subsampled to 15000000.
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
Jensen-Shannon distance (filtered/deduped BAM)
rep1
rep2
rep3
AUC
0.24710892583071029
0.2687381569695909
0.21885365118214736
Synthetic AUC
0.49320614723692013
0.49372862409008295
0.4897345001503545
X-intercept
0.04953966245904452
0.04476867390063733
0.09646136716458414
Synthetic X-intercept
1.9553127716174093e-185
7.1262433716555e-218
1.6398375985467922e-80
Elbow Point
0.7096441413218292
0.6970152887136489
0.7014251108441717
Synthetic Elbow Point
0.5055645379760925
0.5077646579035788
0.4915156256548443
JS Distance
0.25120883136124034
0.21824202617793453
0.2840662795634995
Synthetic JS Distance
0.3565146322406388
0.3260932549747285
0.3765172862657871
% Genome Enriched
24.898933262031232
25.268679886821218
24.410061577878256
Diff. Enrichment
30.59694709332332
26.92947227033608
34.82361033172099
CHANCE Divergence
0.2630576103680777
0.23179139631161688
0.29941245645630304
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for spp raw peaks
rep1
rep2
rep3
rep1-pr1
rep2-pr1
rep3-pr1
rep1-pr2
rep2-pr2
rep3-pr2
pooled
pooled-pr1
pooled-pr2
Fraction of Reads in Peaks
0.4282750000460211
0.4159924783809941
0.42548611338407427
0.4161072342944746
0.40315476183004306
0.3806926139529887
0.41628941617233844
0.4044345165991537
0.38365747913208265
0.4214190224289127
0.4229127920433456
0.4226736811895407
FRiP for overlap peaks
rep1_vs_rep2
rep1_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.3295062357666139
0.29108448041480434
0.2920165046666054
0.2959959795960819
0.2834091938988523
0.2650462467573558
0.3402497464138492
FRiP for IDR peaks
rep1_vs_rep2
rep1_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.16194682556373935
0.14388032763499833
0.1406141549280289
0.12315064181035991
0.11231589795603267
0.10308566287578376
0.17699129264941285
For spp raw peaks:
repX: Peak from true replicate X
repX-prY: Peak from Yth pseudoreplicates from replicate X
pooled: Peak from pooled true replicates (pool of rep1, rep2, ...)
pooled-pr1: Peak from 1st pooled pseudo replicate (pool of rep1-pr1, rep2-pr1, ...)
pooled-pr2: Peak from 2nd pooled pseudo replicate (pool of rep1-pr2, rep2-pr2, ...)
For overlap/IDR peaks:
repX_vs_repY: Comparing two peaks from true replicates X and Y
repX-pr1_vs_repX-pr2: Comparing two peaks from both pseudoreplicates from replicate X
pooled-pr1_vs_pooled-pr2: Comparing two peaks from 1st and 2nd pooled pseudo replicates