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
2060992
1793741
3425822
6403928
Distinct Fragments
1189796
1092847
1936244
5931481
Positions with Two Read
184008
170414
301830
346346
NRF = Distinct/Total
0.577293
0.609256
0.565191
0.926225
PBC1 = OneRead/Distinct
0.719116
0.733604
0.732234
0.934507
PBC2 = OneRead/TwoRead
4.649803
4.704525
4.69729
16.004253
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
15027
3464
N1
6835
2471
N2
6478
2465
N3
11783
2819
Np
20048
3902
N optimal
20048
3902
N conservative
15027
3464
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep2_vs_rep3
rep1_vs_rep3
Rescue Ratio
1.334131895920676
1.1264434180138567
Self Consistency Ratio
1.8189255943192344
1.1436105476673428
Reproducibility Test
pass
pass
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
24595
24618
34148
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
170.0
164.0
171.0
179.0
179.0
25 percentile
680.0
656.0
684.0
638.25
716.0
50 percentile (median)
680.0
656.0
684.0
716.0
716.0
75 percentile
680.0
656.0
684.0
1154.0
716.0
Max size
3778.0
3142.0
3605.0
4353.0
4353.0
Mean
696.416304126855
665.7124461775936
692.2422396626449
930.2314197847257
755.7633679169992
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
2060992
1793741
3425822
Estimated Fragment Length
240
215
160
Cross-correlation at Estimated Fragment Length
0.498252104376504
0.462672973232646
0.47511002182176
Phantom Peak
35
35
35
Cross-correlation at Phantom Peak
0.4152736
0.3850096
0.4488796
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.1592946
0.1627707
0.2422846
NSC (Normalized Strand Cross-correlation coeff.)
3.127865
2.842483
1.960959
RSC (Relative Strand Cross-correlation coeff.)
1.324162
1.349459
1.126965
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
0.1535890182048413
0.16490603466738757
0.21966087066496945
Synthetic AUC
0.4897546393447963
0.48931031190691376
0.4919664079779163
X-intercept
0.11931141398361136
0.1217455671477112
0.05460084876127542
Synthetic X-intercept
7.845036838666058e-81
3.6410682939567845e-74
3.2084187314930106e-132
Elbow Point
0.8417102120706064
0.8306906809842758
0.7934800608937332
Synthetic Elbow Point
0.4990311407857957
0.5121277852373388
0.5035496739923169
JS Distance
0.46290634331898056
0.4345788084286821
0.35343018264185755
Synthetic JS Distance
0.5249415999305265
0.4991455958807507
0.4297920512460509
% Genome Enriched
11.535890790321647
12.837963212363903
13.949692837803596
Diff. Enrichment
49.54685343473578
46.668076080263674
36.55276682721434
CHANCE Divergence
0.4466770304271998
0.41900186111987286
0.33179351016004144
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
0.5713612536574199
0.5359940497136243
0.50107717574851
0.5095473070862457
0.4736183600356435
0.46035861632207
0.5120120446292952
0.474752177257592
0.459018466466944
0.5871043694687397
0.554473945009321
0.5521529345425674
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
0.4470929692557283
0.46321993311686444
0.4634441184471759
0.45934352759921193
0.41106531403127355
0.36902020957506687
0.4901509325968299
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
0.37206537630839603
0.37668250972530576
0.37413679218373486
0.4044082875337753
0.3627792492855383
0.2849799394972152
0.38439721963043133
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