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
1292537
1651374
1031405
2102567
7473669
Distinct Fragments
943943
1453756
886333
1900437
7058120
Positions with Two Read
202959
156789
111573
165190
315475
NRF = Distinct/Total
0.730302
0.880331
0.859345
0.903865
0.944398
PBC1 = OneRead/Distinct
0.717021
0.879152
0.85654
0.904545
0.951129
PBC2 = OneRead/TwoRead
3.334797
8.151541
6.804334
10.406387
21.279604
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
6076
625
N1
3275
177
N2
6421
434
N3
3323
50
N4
7977
105
Np
5769
587
N optimal
6076
625
N conservative
6076
625
Optimal Set
rep2_vs_rep4
rep1_vs_rep2
Conservative Set
rep2_vs_rep4
rep1_vs_rep2
Rescue Ratio
1.0532154619518115
1.0647359454855196
Self Consistency Ratio
2.435725190839695
8.68
Reproducibility Test
borderline
borderline
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
30082
28306
31560
25659
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
112.0
109.0
183.0
202.0
112.0
112.0
25 percentile
416.0
436.0
560.0
600.0
450.0
450.0
50 percentile (median)
416.0
436.0
560.0
600.0
450.0
450.0
75 percentile
416.0
436.0
560.0
600.0
450.0
450.0
Max size
3936.0
568.0
736.0
1337.0
10621.0
10621.0
Mean
418.63991755867295
435.63664947360985
559.8685994930291
600.0666043103785
768.3696
482.66227781435157
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
1292537
1651374
1031405
2102567
Estimated Fragment Length
135
140
125
110
Cross-correlation at Estimated Fragment Length
0.254241782622474
0.359732744663143
0.257960195915871
0.420740695502499
Phantom Peak
30
30
35
30
Cross-correlation at Phantom Peak
0.2494121
0.3548259
0.255608
0.4192149
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.2389925
0.3439556
0.2443822
0.4122605
NSC (Normalized Strand Cross-correlation coeff.)
1.063807
1.04587
1.055561
1.02057
RSC (Relative Strand Cross-correlation coeff.)
1.463518
1.451399
1.209534
1.219397
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
0.3078242177039236
0.3226041664213019
0.3214789872374544
0.3530480043322487
Synthetic AUC
0.48855570733938314
0.49075438948286726
0.48815168644114343
0.49190732285331773
X-intercept
0.07005239259517988
0.05294146998652762
0.0687450726011676
0.04557257621875156
Synthetic X-intercept
1.4094214599912247e-64
1.5946771510762778e-99
3.4544095409581584e-60
2.8172971661942523e-130
Elbow Point
0.5545691332767826
0.5809710094306672
0.5515752706950751
0.527051544334115
Synthetic Elbow Point
0.5198384918508095
0.4927423932083293
0.4929424461922288
0.5147748565062529
JS Distance
0.12450935438274609
0.10674333020754954
0.09387188189525088
0.055001659841304354
Synthetic JS Distance
0.2389304885422796
0.22959183486271864
0.2130332913711937
0.18959424067920874
% Genome Enriched
34.24958834389501
32.112968414749766
41.97056035127988
37.06022653560201
Diff. Enrichment
19.470747057803223
17.299622596078674
18.555001343673727
12.68444110589213
CHANCE Divergence
0.16591288523770692
0.14758220377158685
0.15998890343444133
0.10787826545274183
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
0.26861212158451026
0.24867335708759658
0.2706301594226021
0.21808952199178006
0.19229303223629535
0.23856008847918234
0.1922440973872529
0.26205520521685227
0.1936972777071921
0.2507191657677095
0.19898560297044726
0.255155602027804
0.14908222792845405
0.17718507450043366
0.17843188131505847
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
0.07914824219208336
0.0629133999930935
0.07038615129459579
0.06797212141770882
0.07606550333562723
0.06372672563643256
0.06746988453699722
0.09065302430998223
0.04203628931035137
0.08158379844690244
0.07496351672621451
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
0.024940460098488146
0.016372590525166448
0.018271495082556464
0.01645882631939334
0.019156175122003617
0.015518627217733743
0.023857007917968884
0.0198615061074608
0.0072029695513875424
0.008076462114093137
0.02427957339228911
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