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
5322268
1452737
2714178
2236621
5524677
Distinct Fragments
4955242
1417685
2613389
2179336
5291200
Positions with Two Read
285934
28330
86940
49496
185082
NRF = Distinct/Total
0.93104
0.975872
0.962866
0.974388
0.957739
PBC1 = OneRead/Distinct
0.935547
0.978516
0.964699
0.976044
0.962032
PBC2 = OneRead/TwoRead
16.213049
48.966749
28.998539
42.975735
27.502966
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
2506
65
N1
7602
69
N2
5472
48
N3
7822
16
N4
8269
5
Np
2777
136
N optimal
2777
136
N conservative
2506
65
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep3
rep2_vs_rep3
Rescue Ratio
1.1081404628890663
2.0923076923076924
Self Consistency Ratio
1.5111476608187135
13.8
Reproducibility Test
pass
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
17599
46253
30626
31701
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
125.0
129.0
476.0
524.0
149.0
149.0
25 percentile
490.0
400.0
476.0
524.0
410.0
410.0
50 percentile (median)
490.0
400.0
476.0
524.0
410.0
410.0
75 percentile
490.0
400.0
476.0
524.0
410.0
410.0
Max size
490.0
400.0
476.0
524.0
410.0
410.0
Mean
489.9792601852378
399.9895574341124
476.0
524.0
406.5735294117647
409.8321930140439
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
5322268
1452737
2714178
2236621
Estimated Fragment Length
130
105
110
130
Cross-correlation at Estimated Fragment Length
0.656092713610721
0.371766278318909
0.509811195447864
0.467779394761086
Phantom Peak
30
35
30
30
Cross-correlation at Phantom Peak
0.6558685
0.3692765
0.5094356
0.4675732
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.650976
0.3597197
0.5037854
0.4617572
NSC (Normalized Strand Cross-correlation coeff.)
1.00786
1.033489
1.011961
1.013042
RSC (Relative Strand Cross-correlation coeff.)
1.045818
1.260523
1.066472
1.035451
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
0.3909588865503199
0.3318325424131577
0.37518602124767014
0.3713798588574375
Synthetic AUC
0.4949742011274266
0.49059104122113134
0.49307908455701144
0.49241774328242194
X-intercept
0.036189126899400376
0.05820870207225454
0.040954214847998084
0.04322900557722815
Synthetic X-intercept
0.0
4.684337571294189e-96
1.2567621958820313e-178
1.3828003481933983e-148
Elbow Point
0.4564547186941903
0.5432140398487464
0.48035997565574834
0.48316156002753696
Synthetic Elbow Point
0.5036571274951887
0.4931514690223175
0.5000553115218107
0.5076740893748594
JS Distance
0.018759691219164137
0.10392723790255061
0.04138728893510009
0.04644946559410251
Synthetic JS Distance
0.1509176838871308
0.20605918463783388
0.15971107485643374
0.16088891587951143
% Genome Enriched
39.013259635434856
41.41296431172615
38.99609893344242
42.12992247752646
Diff. Enrichment
7.4044435476696036
17.36001784935483
10.17832375134527
10.847099939711601
CHANCE Divergence
0.0629241188802819
0.1490057639367185
0.08652358530388723
0.0925263167680375
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
0.12129098116212565
0.306742966047997
0.20190537768030908
0.21981283533973836
0.20841756052267604
0.21811138724469586
0.24455330905533626
0.2632183433932344
0.21266559307225322
0.2096552287996301
0.24483474390387433
0.25875280944192475
0.03483139203479517
0.1252804236268343
0.11741816035204465
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
0.02077984129910165
0.020791826555804832
0.018917654399398662
0.019844275738540015
0.018104177461223895
0.01814013323133343
0.05555609289009682
0.0501107628504994
0.055845656077456264
0.060838368251212575
0.022702222808963148
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
0.0008808269254695023
0.0011566667140710401
0.0009876209292276853
0.0012231222792237454
0.0007089189897716567
0.000906228514303107
0.0012599823324402834
0.0013328192828149482
0.0003870979082574136
0.00015674203881061208
0.0023898959634997058
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