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
ctl2
Total Fragments
1818466
7865547
5069622
12797571
11751540
Distinct Fragments
1716158
6456309
4288346
12676706
11624394
Positions with Two Read
87660
907975
575364
112813
117699
NRF = Distinct/Total
0.943739
0.820834
0.845891
0.990556
0.98918
PBC1 = OneRead/Distinct
0.945088
0.826825
0.843635
0.990925
0.989612
PBC2 = OneRead/TwoRead
18.502396
5.879281
6.28784
111.349419
97.737797
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
16345
156384
92779
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
70.0
76.0
74.0
79.0
79.0
25 percentile
280.0
304.0
296.0
316.0
316.0
50 percentile (median)
280.0
304.0
296.0
316.0
316.0
75 percentile
280.0
304.0
296.0
316.0
316.0
Max size
481.0
692.0
318.0
767.0
767.0
Mean
277.7061486693178
303.2199905361162
295.651591416161
285.83435700575814
304.829892885011
rep1rep2rep3idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
1818466
7865547
5069622
Estimated Fragment Length
105
100
90
Cross-correlation at Estimated Fragment Length
0.0456593603666933
0.106524928827216
0.0639911888505514
Phantom Peak
40
40
35
Cross-correlation at Phantom Peak
0.03910966
0.09675796
0.05777459
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.02201109
0.06788273
0.04957696
NSC (Normalized Strand Cross-correlation coeff.)
2.07438
1.569249
1.290745
RSC (Relative Strand Cross-correlation coeff.)
1.383055
1.338247
1.75834
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
0.08601218004100818
0.17802645543884493
0.15149868485496332
Synthetic AUC
0.45394244184844057
0.47632874793774593
0.4709680772830374
X-intercept
0.7731413170827285
0.44120951843987793
0.5580527264948038
Synthetic X-intercept
0.0007819167889891444
1.2281704150981013e-14
1.223332523162236e-09
Elbow Point
0.7792812786154997
0.7266067665535042
0.5824906131164453
Synthetic Elbow Point
0.5210879635686168
0.5023836964312945
0.4948988238805788
JS Distance
0.1799700681348484
0.1205227340070402
0.11551036227486353
Synthetic JS Distance
0.17010955279624898
0.24065570097694966
0.18961652043167432
% Genome Enriched
21.903727962316513
30.424698999932023
41.359068789161995
Diff. Enrichment
67.73664247779159
33.13700119206645
41.84216531505146
CHANCE Divergence
0.6741085868675498
0.29189515912240466
0.4643960025626234
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
0.060628695451698016
0.11822825945121626
0.08524419012920344
0.04475295345040788
0.07800803670383694
0.04935957076852439
0.046555414018672
0.07792199751159301
0.04957144839545772
0.1425285916884251
0.10342834859196033
0.10416660319594206
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
0.03119600562593768
0.027288748132658025
0.03235162703782098
0.02953974378977814
0.035317851531161036
0.0166410085188776
0.03912781450215879
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
0.02583953121568823
0.02213551364742066
0.028114295078555236
0.019861415467160984
0.029168348074774173
0.01350847929606336
0.031442218568624905
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