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
4621582
3159093
3994089
17830145
Distinct Fragments
3745651
2633078
3095432
15961259
Positions with Two Read
597517
377743
569796
1390649
NRF = Distinct/Total
0.810469
0.833492
0.775003
0.895184
PBC1 = OneRead/Distinct
0.807056
0.830876
0.769046
0.900976
PBC2 = OneRead/TwoRead
5.059185
5.791665
4.177865
10.341009
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
18455
3998
N1
12991
2482
N2
10673
1834
N3
11944
2121
Np
25166
5093
N optimal
25166
5093
N conservative
18455
3998
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep3
rep1_vs_rep3
Rescue Ratio
1.3636412896234082
1.273886943471736
Self Consistency Ratio
1.217183547268809
1.3533260632497273
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
42290
29082
37299
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
110.0
109.0
111.0
110.0
110.0
25 percentile
440.0
436.0
444.0
419.0
440.0
50 percentile (median)
440.0
436.0
444.0
440.0
440.0
75 percentile
440.0
436.0
444.0
440.0
440.0
Max size
10683.0
10624.0
13359.0
13386.0
13386.0
Mean
441.57786710806334
440.2834743140087
453.1131933832006
480.5383860200275
447.9873639036796
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
4621582
3159093
3994089
Estimated Fragment Length
145
175
160
Cross-correlation at Estimated Fragment Length
0.582748276931258
0.503172849376809
0.534979509989116
Phantom Peak
35
40
30
Cross-correlation at Phantom Peak
0.5731649
0.4926747
0.5272721
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.5535082
0.4724178
0.5057254
NSC (Normalized Strand Cross-correlation coeff.)
1.052827
1.065101
1.057846
RSC (Relative Strand Cross-correlation coeff.)
1.487537
1.518252
1.357709
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
0.327876277938834
0.31588877893401157
0.3233072640124418
Synthetic AUC
0.4942349398974187
0.49312958988674277
0.49366181874345305
X-intercept
0.03951347140945495
0.04336216021889792
0.041446803411292414
Synthetic X-intercept
3.860584806093472e-258
2.7901981681864027e-181
2.792481771444423e-213
Elbow Point
0.6306357227026703
0.6409754538736544
0.6295112744412711
Synthetic Elbow Point
0.5090187648102765
0.4976308925625949
0.500630156624896
JS Distance
0.14560035313290576
0.16382696422415716
0.15326302993246338
Synthetic JS Distance
0.24329420108013933
0.2567409322323278
0.24837895467149593
% Genome Enriched
27.47069044718289
26.928438754518766
27.896902274860693
Diff. Enrichment
16.54329918356252
17.97514038013457
17.307828286325673
CHANCE Divergence
0.14286659133575824
0.15505957546368399
0.14914187280210617
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
0.33046072582471764
0.2867897336351716
0.3175221597450315
0.29094201687759685
0.29792821995580104
0.29726108884666763
0.2934939723247301
0.2965700964698016
0.2961406018835463
0.3479205290465809
0.35378504201039856
0.34265915634353383
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
0.1835928772328567
0.19804616954391385
0.17663234444885204
0.16968169779564304
0.1665097363910203
0.16712723296129064
0.23327390256909128
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
0.08823975824901696
0.09234652496579178
0.08449070740067519
0.06946055661378693
0.06528613201946111
0.0682872061926776
0.10740411093570405
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