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
4679763
2844414
1233383
16001732
Distinct Fragments
2754282
1925925
873445
13721629
Positions with Two Read
694781
407628
173540
1079415
NRF = Distinct/Total
0.588552
0.67709
0.70817
0.857509
PBC1 = OneRead/Distinct
0.611204
0.711953
0.738261
0.912407
PBC2 = OneRead/TwoRead
2.422961
3.363773
3.715743
11.598613
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
13346
5893
N1
11100
4828
N2
9702
4021
N3
5881
1869
Np
14437
6271
N optimal
14437
6271
N conservative
13346
5893
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep2
rep1_vs_rep2
Rescue Ratio
1.0817473400269744
1.0641438995418293
Self Consistency Ratio
1.8874341098452645
2.583199571963617
Reproducibility Test
pass
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
Number of peaks
29995
30343
22895
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
96.0
98.0
100.0
94.0
94.0
25 percentile
384.0
390.0
400.0
218.0
376.0
50 percentile (median)
384.0
390.0
400.0
376.0
376.0
75 percentile
384.0
390.0
400.0
376.0
376.0
Max size
3383.0
10132.0
6312.0
10705.0
10705.0
Mean
376.2109684947491
390.4693009919916
400.72710198733347
350.57327380003187
364.7288910438457
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
4679763
2844414
1233383
Estimated Fragment Length
145
145
135
Cross-correlation at Estimated Fragment Length
0.501905516579447
0.431028751491673
0.299349265787419
Phantom Peak
40
40
40
Cross-correlation at Phantom Peak
0.4741423
0.4072397
0.2809193
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.4331683
0.3657361
0.2416185
NSC (Normalized Strand Cross-correlation coeff.)
1.158685
1.178524
1.238933
RSC (Relative Strand Cross-correlation coeff.)
1.677582
1.573178
1.468948
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
0.28875918637404246
0.2718140545000659
0.23080384258784753
Synthetic AUC
0.49327374745725133
0.49196595485812744
0.48808564949900946
X-intercept
0.04376935007695117
0.050554228122810484
0.10838438460018325
Synthetic X-intercept
3.3340796137695087e-189
3.325981581710734e-132
1.653802133660937e-59
Elbow Point
0.6999902189166153
0.6991438557829158
0.7041841079352532
Synthetic Elbow Point
0.5112760633858362
0.50436982368292
0.5123382611941688
JS Distance
0.2067922740739427
0.22754806082233808
0.271446533707158
Synthetic JS Distance
0.30447244360366077
0.32303276234270356
0.35458868063785953
% Genome Enriched
21.561500058886114
23.507935652448165
27.603814223292154
Diff. Enrichment
22.37554119520082
24.986419421926676
31.714867023482185
CHANCE Divergence
0.19622173821399938
0.21622409496925854
0.2717296739769245
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
0.3207768768474044
0.34443381620090774
0.3602190383011991
0.326767590753741
0.32992806643820816
0.305325021296018
0.32727634860520965
0.3296659871219239
0.3052143045974544
0.36357594217344574
0.32998693942098456
0.3305295518473387
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
0.2338861908012722
0.21541137884846528
0.21206266293360942
0.2148080287040458
0.2195693348817752
0.21353071032670465
0.2405209446975272
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
0.17337859239647024
0.1583703523017712
0.15477788886791688
0.15057067476445593
0.14768101616756515
0.11875043778286422
0.17868282412125447
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