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
1198083
288264
620662
421854
7383487
Distinct Fragments
1130980
279646
600469
414281
7180147
Positions with Two Read
59697
8062
18720
7151
171568
NRF = Distinct/Total
0.943991
0.970104
0.967465
0.982048
0.97246
PBC1 = OneRead/Distinct
0.944199
0.970348
0.967689
0.982258
0.974883
PBC2 = OneRead/TwoRead
17.888169
33.658397
31.039904
56.905468
40.799007
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
4909
211
N1
4315
193
N2
306
31
N3
1523
57
N4
931
38
Np
6497
458
N optimal
6497
458
N conservative
4909
211
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep3
rep1_vs_rep3
Rescue Ratio
1.323487471990222
2.170616113744076
Self Consistency Ratio
14.101307189542483
6.225806451612903
Reproducibility Test
borderline
fail
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
27049
7847
19643
13587
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
129.0
262.0
284.0
187.0
129.0
129.0
25 percentile
500.0
490.0
490.0
510.0
516.0
516.0
50 percentile (median)
500.0
490.0
490.0
510.0
516.0
516.0
75 percentile
500.0
490.0
490.0
510.0
516.0
516.0
Max size
1123.0
3037.0
2767.0
1358.0
6347.0
6347.0
Mean
500.11882139820324
495.6681534344335
491.26497989105536
510.21579450945757
706.3471615720524
529.3546252116362
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
1198083
288264
620662
421854
Estimated Fragment Length
120
105
125
120
Cross-correlation at Estimated Fragment Length
0.320504414452819
0.107473746879476
0.203380859524224
0.153892904421686
Phantom Peak
30
30
35
35
Cross-correlation at Phantom Peak
0.3193235
0.107116
0.2024023
0.1535106
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.3057926
0.1011496
0.1938662
0.1447977
NSC (Normalized Strand Cross-correlation coeff.)
1.04811
1.062522
1.049078
1.062813
RSC (Relative Strand Cross-correlation coeff.)
1.087274
1.059968
1.114643
1.043876
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
0.3210469125751732
0.2328805471723006
0.28607985051499346
0.2589958591176447
Synthetic AUC
0.4895150849115998
0.4791132773259738
0.4856680826863349
0.4827049163996491
X-intercept
0.057820400767852366
0.28740386879462343
0.113397906365113
0.18789036153714148
Synthetic X-intercept
3.9292802935931675e-77
6.446502692251736e-19
7.33625638142448e-41
7.738008451596911e-28
Elbow Point
0.5572736451835623
0.611889339862473
0.6325303608985948
0.6555759092305912
Synthetic Elbow Point
0.515743296868069
0.4962882630385415
0.4865736547973779
0.5012964203784969
JS Distance
0.12866641005633314
0.1791832439667173
0.1656958324350119
0.1745683559016435
Synthetic JS Distance
0.22556510566688506
0.23125470329635942
0.24374365042953264
0.24022644659552497
% Genome Enriched
34.806580223411515
38.2042471335241
34.65173265647387
33.24533362067933
Diff. Enrichment
18.009887635006383
34.29661296276543
23.859439178019993
28.71025497508713
CHANCE Divergence
0.15338033893104916
0.30187904794618586
0.20387238776067973
0.24600815107132804
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
0.2500908522288498
0.14521306185064767
0.22288311167546615
0.1876924900717207
0.19913236121448463
0.09444966554259494
0.16407599872205306
0.10840351276506961
0.21832838887549402
0.10419387607023654
0.13477494042204988
0.1408384585109561
0.18968152953939768
0.23232202154369483
0.23674796326548134
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
0.05250350591662661
0.08623886722143098
0.07565247710270968
0.04614932309385652
0.04252090692586085
0.06333408520379671
0.0693254920871203
0.03684226892817779
0.049191257133449504
0.035573469266789164
0.10195646953487025
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
0.019770008629637115
0.024676064642501792
0.02438935497208428
0.019996379682975237
0.01931362190850641
0.022047487706512967
0.016718557266605864
0.03269954357462952
0.02186368652432097
0.01303775709797878
0.03214144991672056
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