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
2400052
1040804
1382396
876672
8092692
Distinct Fragments
2257463
1018611
1289534
824750
7720272
Positions with Two Read
119766
18650
79741
45470
295598
NRF = Distinct/Total
0.940589
0.978677
0.932825
0.940774
0.953981
PBC1 = OneRead/Distinct
0.942956
0.980312
0.933565
0.941283
0.958853
PBC2 = OneRead/TwoRead
17.773734
53.54193
15.097177
17.073301
25.042805
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
5070
423
N1
7996
611
N2
4011
130
N3
5742
129
N4
3255
42
Np
4976
534
N optimal
5070
534
N conservative
5070
423
Optimal Set
rep1_vs_rep3
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep3
rep1_vs_rep2
Rescue Ratio
1.0188906752411575
1.2624113475177305
Self Consistency Ratio
2.4565284178187405
14.547619047619047
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
24244
25478
24696
26337
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
111.0
149.0
145.0
194.0
100.0
100.0
25 percentile
444.0
596.0
580.0
660.0
400.0
400.0
50 percentile (median)
444.0
596.0
580.0
660.0
400.0
400.0
75 percentile
444.0
596.0
580.0
660.0
400.0
400.0
Max size
444.0
596.0
10627.0
3702.0
1987.0
1987.0
Mean
443.728757630754
595.7944893633723
587.8760123096857
661.3694042601663
392.71535580524346
399.2420118343195
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
2400052
1040804
1382396
876672
Estimated Fragment Length
125
130
110
125
Cross-correlation at Estimated Fragment Length
0.471391778730553
0.306251630392057
0.345031452020133
0.257257811398507
Phantom Peak
35
35
35
35
Cross-correlation at Phantom Peak
0.4679734
0.3058346
0.344031
0.2565063
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.4589817
0.2911571
0.3317279
0.2450143
NSC (Normalized Strand Cross-correlation coeff.)
1.027038
1.051843
1.040104
1.04997
RSC (Relative Strand Cross-correlation coeff.)
1.380166
1.028411
1.081318
1.065396
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
0.3540455402416667
0.32540969132091757
0.33435731381787454
0.31989310191201553
Synthetic AUC
0.49256368015874774
0.4889169053882003
0.490184516110064
0.48772574323226353
X-intercept
0.0431761796505907
0.061684533658538536
0.05339539376907371
0.07279992495264631
Synthetic X-intercept
1.5534345969780088e-154
6.312228590611763e-69
3.59219787735787e-88
5.125269778798024e-56
Elbow Point
0.5454904723791807
0.5818465241698385
0.5501050862545582
0.5793496029076858
Synthetic Elbow Point
0.513031933529281
0.5167933468525627
0.49950090662922036
0.5161015425791736
JS Distance
0.08775631469496563
0.11882636531125837
0.11170988026917077
0.12333948960369603
Synthetic JS Distance
0.191682272071672
0.21099600058194307
0.2094585682870925
0.21305514014049784
% Genome Enriched
36.20715464744709
38.31446837133271
35.749485546398624
39.26513471598768
Diff. Enrichment
12.827394295795486
17.70254662392876
16.407434318837154
19.19275840417829
CHANCE Divergence
0.10911926069459886
0.15111737785425094
0.139669628253619
0.1642684342323924
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
0.19878626484443315
0.23968393498984217
0.2293312487546175
0.252516814667656
0.24954426532396828
0.21676043131739334
0.24235373461473614
0.20850905957538476
0.24866629291134243
0.19314150648538833
0.23889731917046028
0.1922233658057876
0.13084141220193835
0.15959532525053818
0.1661698358801893
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
0.060580471126634054
0.06014628102094125
0.050424030167653705
0.049361469981064235
0.041957599191916814
0.04305034470166594
0.08996925333558997
0.05834407719956243
0.07577060437454974
0.04914071662797099
0.06081330304003016
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
0.01640903616054469
0.01368793971181115
0.009657094728906348
0.011439317193265582
0.008669261622821871
0.008638340072268082
0.019337871287128713
0.008181942491014222
0.015642770650357903
0.01314655688264439
0.018470472864130706
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