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
rep5
ctl1
Total Fragments
3208678
3913842
1564984
930622
1094546
15759633
Distinct Fragments
2984858
3656142
1342295
601079
1020730
14663381
Positions with Two Read
184273
213278
170593
152244
63695
875459
NRF = Distinct/Total
0.930245
0.934157
0.857705
0.64589
0.93256
0.930439
PBC1 = OneRead/Distinct
0.932962
0.936925
0.855328
0.622061
0.933137
0.936159
PBC2 = OneRead/TwoRead
15.112143
16.061347
6.730065
2.455979
14.95378
15.680075
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
2219
213
N1
5330
80
N2
6356
66
N3
4966
268
N4
999
59
N5
5065
33
Np
2622
139
N optimal
2622
213
N conservative
2219
213
Optimal Set
pooled-pr1_vs_pooled-pr2
rep3_vs_rep4
Conservative Set
rep1_vs_rep3
rep3_vs_rep4
Rescue Ratio
1.181613339342046
1.5323741007194245
Self Consistency Ratio
6.362362362362362
8.121212121212121
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
rep5
Number of peaks
18018
19755
30174
19827
25209
Top 300000 raw peaks from spp with FDR 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
rep4
rep5
idr_opt
overlap_opt
Min size
198.0
151.0
120.0
99.0
800.0
128.0
126.0
25 percentile
684.0
604.0
470.0
390.0
800.0
504.0
504.0
50 percentile (median)
684.0
604.0
470.0
390.0
800.0
504.0
504.0
75 percentile
684.0
604.0
470.0
390.0
800.0
504.0
504.0
Max size
10679.0
10661.0
2733.0
965.0
10695.0
10648.0
10648.0
Mean
692.9543234543235
612.6592761326247
470.49423344601314
390.12170272860243
808.4862549089611
1399.9483568075118
573.8318077803203
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
rep5
Number of Subsampled Reads
3208678
3913842
1564984
930622
1094546
Estimated Fragment Length
150
105
150
135
150
Cross-correlation at Estimated Fragment Length
0.534303542568179
0.584701689189796
0.339603019310921
0.180950924991158
0.292868784924775
Phantom Peak
35
30
40
30
30
Cross-correlation at Phantom Peak
0.5344821
0.5852492
0.3353381
0.1754299
0.2938288
Argmin of Cross-correlation
1500
1500
1500
1500
1500
Minimum of Cross-correlation
0.5298156
0.5810422
0.3235209
0.1649816
0.2826909
NSC (Normalized Strand Cross-correlation coeff.)
1.008471
1.006298
1.04971
1.096795
1.036004
RSC (Relative Strand Cross-correlation coeff.)
0.9617302
0.869861
1.360911
1.528415
0.9138073
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
rep5
AUC
0.3738907035305189
0.3845473302682687
0.32705769206066565
0.28788614458259537
0.33896959497934365
Synthetic AUC
0.493541510195727
0.49416203601970676
0.49036707059895623
0.4856045769572665
0.4889628272903833
X-intercept
0.03994933917184728
0.0380535534495709
0.05386908091349304
0.11713119076918159
0.058335863711185335
Synthetic X-intercept
2.3903039552575013e-205
1.1026321797741657e-251
1.371971671175897e-91
1.707136435521003e-40
1.657908682326754e-69
Elbow Point
0.48996771770571973
0.4720406565131926
0.5507946718233103
0.6263544180397296
0.5533416968980231
Synthetic Elbow Point
0.5061691918721752
0.5073198758320864
0.5020256147505144
0.5212184661295013
0.50596580614627
JS Distance
0.06114603313138906
0.04301788386159794
0.12336145759863436
0.1582800819387885
0.10074520244374248
Synthetic JS Distance
0.17157049332203478
0.1596171510389945
0.2193265061116288
0.23317549314025432
0.19463691833412292
% Genome Enriched
37.758697840921805
38.29507135665543
35.90785961996388
35.71029038083518
40.72523292794017
Diff. Enrichment
8.785506720927067
7.650170186620686
16.39674256338481
24.143514515817294
16.29840187268098
CHANCE Divergence
0.07466656745712881
0.0650131934776388
0.13960966687240128
0.20656396757890513
0.13940253179198764
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for spp raw peaks
rep1
rep2
rep3
rep4
rep5
rep1-pr1
rep2-pr1
rep3-pr1
rep4-pr1
rep5-pr1
rep1-pr2
rep2-pr2
rep3-pr2
rep4-pr2
rep5-pr2
pooled
pooled-pr1
pooled-pr2
Fraction of Reads in Peaks
0.1616784981900296
0.1518641808385076
0.2657174324170706
0.21194374385499937
0.24113380371735113
0.222252661760286
0.2141898360703865
0.22112251661984225
0.12159127155066111
0.24757736732187208
0.22197737603498124
0.22495048622176844
0.22478382763492596
0.1341426922583679
0.24169950443067648
0.07069706863268617
0.11666988575506904
0.1138941708334065
FRiP for overlap peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep5
rep4_vs_rep5
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.025412447853350038
0.029192931501585962
0.02302033809723988
0.021049223132403688
0.026674812509618534
0.02075496058079929
0.021460715587858906
0.029364593263876766
0.024205754490628322
0.019570163904964247
0.056697555409563984
0.0555687846005218
0.07609695387781555
0.02804750928121167
0.05899657867308311
0.03250552839700759
FRiP for IDR peaks
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep4
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep5
rep4_vs_rep5
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.009329951052641908
0.00869887201076656
0.008860101860908527
0.008280149517188383
0.008434769046760788
0.008461313615297934
0.008188844464321311
0.011339385219529778
0.008562844007796047
0.008531754843867327
0.010623103254754995
0.007561214687284405
0.017070862607827825
0.010164118786356207
0.011653047228260239
0.010459076434930173
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