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
1792313
1410144
664248
6981057
Distinct Fragments
1608704
1227758
601489
6517772
Positions with Two Read
148393
139488
52405
359890
NRF = Distinct/Total
0.897558
0.870661
0.905519
0.933637
PBC1 = OneRead/Distinct
0.897709
0.870908
0.904856
0.939456
PBC2 = OneRead/TwoRead
9.731915
7.665634
10.385669
17.013982
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
8101
304
N1
8133
89
N2
5796
225
N3
2306
53
Np
7531
406
N optimal
8101
406
N conservative
8101
304
Optimal Set
rep1_vs_rep2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep2
rep1_vs_rep2
Rescue Ratio
1.0756871597397424
1.3355263157894737
Self Consistency Ratio
3.5268863833477884
4.245283018867925
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
37432
26110
22262
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
338.0
155.0
305.0
139.0
130.0
25 percentile
490.0
510.0
570.0
496.0
496.0
50 percentile (median)
490.0
510.0
570.0
496.0
496.0
75 percentile
490.0
510.0
570.0
496.0
496.0
Max size
2821.0
2976.0
3706.0
3886.0
3886.0
Mean
490.907645864501
510.83416315587897
572.4073308777289
642.3029556650247
503.28712504629056
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
1792313
1410144
664248
Estimated Fragment Length
135
135
155
Cross-correlation at Estimated Fragment Length
0.379845034085087
0.322833985021719
0.198252965590054
Phantom Peak
35
35
35
Cross-correlation at Phantom Peak
0.3781771
0.3193265
0.1959097
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.3715398
0.3074306
0.187103
NSC (Normalized Strand Cross-correlation coeff.)
1.022353
1.050103
1.059592
RSC (Relative Strand Cross-correlation coeff.)
1.251293
1.294852
1.266079
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
0.3407340818391625
0.3168973822335345
0.2842632561860546
Synthetic AUC
0.49121623217680377
0.4899445219125029
0.4856763220954588
X-intercept
0.049250172116180914
0.05773125929177933
0.11591351286630813
Synthetic X-intercept
2.1641402572870386e-110
5.869141220750066e-84
6.606515627761278e-41
Elbow Point
0.5544136575436776
0.5887111740817976
0.6331400975823913
Synthetic Elbow Point
0.5148258897944262
0.49236148799143226
0.5227613979208459
JS Distance
0.06079512194427357
0.09858043574335612
0.1295122758940286
Synthetic JS Distance
0.2041837055165246
0.2332884623995027
0.24427808621747937
% Genome Enriched
38.2912106003612
36.51437294832523
34.9588417828243
Diff. Enrichment
13.41862376204157
16.354251103453443
23.87255743905397
CHANCE Divergence
0.11436351596790745
0.13952021924285926
0.20414219306331938
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
0.2771212124006928
0.25161955951841025
0.25645749845241583
0.28274944447655365
0.245154992006742
0.1810445378425048
0.24240773442073005
0.2458160038856562
0.18399635095950218
0.19047672355567954
0.22949811728680333
0.2214644927952236
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
0.09877257627516611
0.08169553886688706
0.07781012750154213
0.08168505398141777
0.08892539733086298
0.05865995503860815
0.09351825778797354
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
0.021056885855420558
0.01722113289478934
0.020419650835395018
0.011818791645069795
0.019298812751900207
0.021081354054670445
0.023224059009111026
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