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
rep4
ctl1
Total Fragments
2340610
1797993
2411042
1260432
5896700
Distinct Fragments
2067082
1743792
2306062
1220305
5728500
Positions with Two Read
194378
48363
91784
36751
145471
NRF = Distinct/Total
0.883138
0.969855
0.956459
0.968164
0.971476
PBC1 = OneRead/Distinct
0.888427
0.970745
0.957665
0.968632
0.973011
PBC2 = OneRead/TwoRead
9.447839
35.001489
24.06122
32.163125
38.316173
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
5037
380
N1
7413
65
N2
6722
13
N3
8364
307
N4
4625
223
Np
4678
366
N optimal
5037
380
N conservative
5037
380
Optimal Set
rep3_vs_rep4
rep3_vs_rep4
Conservative Set
rep3_vs_rep4
rep3_vs_rep4
Rescue Ratio
1.0767421975203078
1.0382513661202186
Self Consistency Ratio
1.8084324324324323
23.615384615384617
Reproducibility Test
pass
borderline
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
rep4
Number of peaks
26160
23539
25738
29156
Top 300000 raw peaks from spp with FDR 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
rep4
idr_opt
overlap_opt
Min size
166.0
624.0
116.0
110.0
111.0
111.0
25 percentile
536.0
624.0
464.0
440.0
424.0
424.0
50 percentile (median)
536.0
624.0
464.0
440.0
424.0
424.0
75 percentile
536.0
624.0
464.0
440.0
424.0
424.0
Max size
536.0
624.0
464.0
440.0
424.0
424.0
Mean
535.9858562691131
624.0
463.8879089284327
439.93754287282206
402.58684210526314
422.33154655548935
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
2340610
1797993
2411042
1260432
Estimated Fragment Length
130
110
120
120
Cross-correlation at Estimated Fragment Length
0.434040495748207
0.413585083935786
0.481988858826195
0.335819420314785
Phantom Peak
40
30
30
30
Cross-correlation at Phantom Peak
0.4321364
0.4134118
0.4805084
0.3343228
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.4260019
0.4071137
0.470807
0.3243789
NSC (Normalized Strand Cross-correlation coeff.)
1.01887
1.015896
1.02375
1.035269
RSC (Relative Strand Cross-correlation coeff.)
1.310402
1.02752
1.152606
1.150507
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
rep4
AUC
0.3625400947361172
0.3570840886313499
0.35192663018776255
0.3240222818586878
Synthetic AUC
0.49222505177919296
0.4915345408278782
0.4926401808525688
0.48988681491293
X-intercept
0.044034895879061675
0.0463595584537901
0.04327065058625396
0.05683949651599927
Synthetic X-intercept
3.0265429033070455e-141
5.972706425184854e-119
8.425547697116372e-158
5.411145339469899e-83
Elbow Point
0.5173022739789443
0.5120363645070917
0.533044130675968
0.5829655111863162
Synthetic Elbow Point
0.506802199732418
0.4955866579281596
0.5016619490297937
0.49897254783481154
JS Distance
0.0531088145475458
0.061351037860976496
0.07539075235638629
0.11562837160865547
Synthetic JS Distance
0.17404968373133411
0.17778510353120594
0.19323831373557432
0.21951078414042935
% Genome Enriched
39.331494887737755
39.737363014518664
37.36520947903613
38.07737434849585
Diff. Enrichment
11.70431791595573
12.60636453094423
13.025491779466646
17.868061707798443
CHANCE Divergence
0.09959288441588839
0.10737331146034965
0.11079176225813543
0.15248473277351868
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for spp raw peaks
rep1
rep2
rep3
rep4
rep1-pr1
rep2-pr1
rep3-pr1
rep4-pr1
rep1-pr2
rep2-pr2
rep3-pr2
rep4-pr2
pooled
pooled-pr1
pooled-pr2
Fraction of Reads in Peaks
0.19976599372192383
0.19655898671818456
0.20412034517608646
0.2455574479384294
0.2557289613610144
0.25063892744731775
0.24957716793213955
0.21437123797577642
0.24653544453000686
0.24811549033810354
0.24995292268090216
0.19574856971576765
0.09709416003873621
0.13954721096382078
0.14001900441509313
FRiP for overlap peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep2_vs_rep3
rep2_vs_rep4
rep3_vs_rep4
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.03281571964885539
0.0418301936159079
0.03955438702771246
0.04033999491912948
0.037965665040353834
0.052610080099706656
0.06414162198733783
0.06233216363178336
0.08493871310477079
0.06719805429374072
0.048180738940964196
FRiP for IDR peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep2_vs_rep3
rep2_vs_rep4
rep3_vs_rep4
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.003937674654924211
0.004113415235229106
0.004032533856185309
0.004003356848644341
0.0040116349763652664
0.009672381560348366
0.0036315307075413055
0.0014446566269981133
0.010067203952767197
0.010666849117354073
0.009634519304051016
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