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
9848431
8006608
8764065
12886118
Distinct Fragments
7781161
5837646
6056035
12614390
Positions with Two Read
1414205
1267974
1444620
249602
NRF = Distinct/Total
0.790091
0.729104
0.691008
0.978913
PBC1 = OneRead/Distinct
0.779634
0.71453
0.671013
0.979644
PBC2 = OneRead/TwoRead
4.289661
3.289635
2.812974
49.509283
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
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
499586
499644
499487
Top 500000 raw peaks from macs2 with p-val threshold 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
idr_opt
overlap_opt
Min size
75.0
65.0
85.0
75.0
75.0
25 percentile
75.0
65.0
85.0
155.0
90.0
50 percentile (median)
76.0
66.0
85.0
219.0
130.0
75 percentile
97.0
81.0
99.0
331.0
201.0
Max size
1461.0
2225.0
1804.0
2531.0
2531.0
Mean
95.8848766778893
84.46107428489084
105.20777918144016
270.62504079900776
172.1328065898966
rep1rep2rep3idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
9848431
8006608
8764065
Estimated Fragment Length
75
65
85
Cross-correlation at Estimated Fragment Length
0.108126991703354
0.10142435709485
0.098490660777091
Phantom Peak
40
40
40
Cross-correlation at Phantom Peak
0.08031187
0.07585692
0.07763095
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.0687848
0.04921914
0.06352286
NSC (Normalized Strand Cross-correlation coeff.)
1.571961
2.060669
1.550476
RSC (Relative Strand Cross-correlation coeff.)
3.413025
1.959819
2.478564
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
rep1rep2rep3
Jensen-Shannon distance (filtered/deduped BAM)
rep1
rep2
rep3
AUC
0.15041957913800746
0.11145627009614463
0.14785149628125852
Synthetic AUC
0.48109459354027906
0.4791077531547503
0.47999732964362646
X-intercept
0.4726590771863741
0.5956542427913932
0.4820439614048136
Synthetic X-intercept
3.3463525335548415e-23
6.590156997027791e-19
1.1381032340685773e-20
Elbow Point
0.6501381541328279
0.7216883994513822
0.6746200261513178
Synthetic Elbow Point
0.4933747317133228
0.49761758393503586
0.489582270064137
JS Distance
0.10262390976596435
0.11998380700032359
0.12130561306029806
Synthetic JS Distance
0.2845270154554525
0.2980781936245424
0.2857082034168842
% Genome Enriched
34.20184498746416
27.427494071968106
31.83123203096572
Diff. Enrichment
41.73595792052307
52.60428315518967
43.15039654293079
CHANCE Divergence
0.3912544411815164
0.4869410311522126
0.39498863365810216
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for macs2 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
0.2982281577538895
0.3955448815021983
0.30731964450618315
0.33427298418632906
0.4296976493018712
0.36651408800996743
0.3338032264461684
0.42944480558523107
0.36655696966744417
0.1039796472784089
0.13259497845725782
0.13234202716350008
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
0.06606827708302093
0.06528298632511698
0.06719496112655211
0.11693960106605976
0.1831504731559218
0.13152950236943897
0.07577030771906747
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
0.028590610159263022
0.03135799832672691
0.03253276751522018
0.021707579367706564
0.02599068090908378
0.02609541520700279
0.05101372892728488
For macs2 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