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
rep5
ctl1
ctl2
ctl3
ctl4
ctl5
Total Fragments
899871
1147650
899242
1164837
1538531
1622980
2105576
3771782
3216376
2033094
Distinct Fragments
894408
1138063
893196
1154751
1528258
1619228
2095316
3753321
3201528
2025906
Positions with Two Read
5376
9385
5932
9900
10122
3655
10057
17329
14186
7012
NRF = Distinct/Total
0.993929
0.991646
0.993277
0.991341
0.993323
0.997688
0.995127
0.995105
0.995384
0.996465
PBC1 = OneRead/Distinct
0.993941
0.991666
0.993296
0.991347
0.993328
0.997713
0.995153
0.995246
0.995472
0.996497
PBC2 = OneRead/TwoRead
165.362537
120.253383
149.563048
115.632222
149.976388
442.004104
207.334295
215.562294
224.66044
287.90773
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
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
rep5
Number of peaks
4697
4784
4992
7181
6414
Top 300000 raw peaks from spp with FDR 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
rep4
rep5
idr_opt
overlap_opt
Min size
180.0
120.0
154.0
86.0
152.0
146.0
146.0
25 percentile
720.0
480.0
616.0
344.0
610.0
405.0
584.0
50 percentile (median)
720.0
480.0
616.0
344.0
610.0
584.0
584.0
75 percentile
720.0
480.0
616.0
344.0
610.0
1421.0
584.0
Max size
3514.0
3445.0
3398.0
3222.0
4544.0
6451.0
6451.0
Mean
709.5537577176922
485.5794314381271
608.6398237179487
343.6763681938449
623.6248830682881
1231.4900117508814
756.8855704697986
rep1rep2rep3rep4rep5idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
rep5
Number of Subsampled Reads
1407743
1368596
1234926
1469190
2133400
Estimated Fragment Length
65
145
90
135
120
Cross-correlation at Estimated Fragment Length
0.818655680519487
0.814148863656557
0.799039218245138
0.822855369226837
0.872696832413182
Phantom Peak
55
50
55
50
50
Cross-correlation at Phantom Peak
0.8187089
0.811766
0.7972784
0.8205827
0.8721706
Argmin of Cross-correlation
1500
1500
1500
1500
1500
Minimum of Cross-correlation
0.7780811
0.7802287
0.7638917
0.7967554
0.8504995
NSC (Normalized Strand Cross-correlation coeff.)
1.052147
1.043475
1.046011
1.032758
1.026099
RSC (Relative Strand Cross-correlation coeff.)
0.9986897
1.075558
1.05274
1.095383
1.024282
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
rep1rep2rep3rep4rep5
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for spp raw peaks
rep1
rep2
rep3
rep4
rep5
rep1-pr1
rep2-pr1
rep3-pr1
rep4-pr1
rep5-pr1
rep1-pr2
rep2-pr2
rep3-pr2
rep4-pr2
rep5-pr2
pooled
pooled-pr1
pooled-pr2
Fraction of Reads in Peaks
0.3652690867770532
0.3324488757489889
0.36404483708558943
0.3727953444802631
0.3850940594707029
0.42240272714486954
0.3958925140586207
0.4299717477503252
0.42797807174600866
0.45049188262843237
0.42479813189683463
0.4005921727916341
0.42670603513210864
0.4305901056280771
0.44047042817336246
0.2557601282165435
0.29352745311104866
0.2914473480357397
FRiP for overlap peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep5
rep4_vs_rep5
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.2170389991986466
0.22024165256878284
0.2238256611165524
0.20054776956637876
0.2180871694417238
0.22576627192591933
0.20024432374677234
0.22571338260172735
0.20155293384382514
0.2094612233995192
0.3137623035176508
0.2767491029678823
0.3088651590587788
0.25038555252726113
0.33153085386114783
0.23937343068293118
FRiP for IDR peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep5
rep4_vs_rep5
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.11191737156085833
0.11259558365239071
0.10388798860297391
0.08214103819784525
0.11632695218591399
0.11369299260974089
0.08583260617932509
0.11045214139435491
0.08366868489003651
0.07323996082272283
0.13515396000812607
0.1279623992209638
0.12010572253979962
0.0897764357560938
0.07434066796614382
0.12828189831715786
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