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
641644
1331395
1700037
8032660
Distinct Fragments
391683
1116693
1273975
7546913
Positions with Two Read
107867
160593
266363
400500
NRF = Distinct/Total
0.610437
0.838739
0.749381
0.939528
PBC1 = OneRead/Distinct
0.57376
0.83394
0.735481
0.942488
PBC2 = OneRead/TwoRead
2.083418
5.798852
3.517696
17.759983
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
10893
2002
N1
461
29
N2
5683
895
N3
6049
994
Np
10402
1849
N optimal
10893
2002
N conservative
10893
2002
Optimal Set
rep2_vs_rep3
rep2_vs_rep3
Conservative Set
rep2_vs_rep3
rep2_vs_rep3
Rescue Ratio
1.0472024610651798
1.0827474310438074
Self Consistency Ratio
13.121475054229935
34.275862068965516
Reproducibility Test
borderline
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
Number of peaks
12942
31061
33758
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
94.0
104.0
100.0
96.0
96.0
25 percentile
376.0
416.0
400.0
384.0
384.0
50 percentile (median)
376.0
416.0
400.0
384.0
384.0
75 percentile
376.0
416.0
400.0
384.0
384.0
Max size
448.0
10609.0
3061.0
10695.0
10695.0
Mean
375.7630196260238
421.8466565789897
400.79984003791697
463.3256743256743
398.51409161847056
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
641644
1331395
1700037
Estimated Fragment Length
160
170
155
Cross-correlation at Estimated Fragment Length
0.124644482199196
0.313706482782219
0.323849542327079
Phantom Peak
35
35
30
Cross-correlation at Phantom Peak
0.1199208
0.3046401
0.3138158
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.1118912
0.2839868
0.2963655
NSC (Normalized Strand Cross-correlation coeff.)
1.11398
1.104652
1.092737
RSC (Relative Strand Cross-correlation coeff.)
1.588283
1.43898
1.574993
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
0.25541904819821387
0.29045774638739486
0.2989415982644545
Synthetic AUC
0.4821973698876707
0.48946938290159697
0.49012013421310463
X-intercept
0.20897699674518014
0.06545959613093516
0.05876436580902529
Synthetic X-intercept
3.072381176086196e-26
1.8641797932843555e-76
5.202081909153059e-87
Elbow Point
0.4994901247847249
0.6112855041777838
0.6160130751562054
Synthetic Elbow Point
0.4881618167791316
0.5028883515773498
0.49072138273388394
JS Distance
0.14166667880014028
0.16070057935507756
0.1483665869452347
Synthetic JS Distance
0.23320554756184664
0.27423812251287666
0.26515505305883696
% Genome Enriched
32.42347381676023
30.150088903878046
34.368782465880265
Diff. Enrichment
29.626973490405355
22.54665913894989
21.303898618759227
CHANCE Divergence
0.2537564174525303
0.19265102038599854
0.18167726059784783
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
0.1751411295380987
0.3223604705650233
0.31977123752250486
0.10973229294728655
0.25351532084459305
0.25314292537461563
0.107168126050631
0.25084646036874314
0.25507512471679944
0.2690192831299529
0.28932344716405395
0.28906278775240757
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
0.07320527495318047
0.07435026391765429
0.15868597907358245
0.01824020874626791
0.1307054599776626
0.12484240789875647
0.15355708016126884
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
0.02984160690860771
0.030657938354049147
0.07141819969344186
0.008691070577313897
0.054701070010464044
0.052353918131194166
0.06859061172797357
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