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
1231191
1359055
871177
5636768
Distinct Fragments
952453
1020815
762093
5390325
Positions with Two Read
182470
207895
82739
214108
NRF = Distinct/Total
0.773603
0.751121
0.874785
0.956279
PBC1 = OneRead/Distinct
0.762832
0.739772
0.87628
0.958015
PBC2 = OneRead/TwoRead
3.981816
3.632459
8.071248
24.118734
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
9417
1019
N1
4766
377
N2
4633
317
N3
4177
296
Np
10027
1152
N optimal
10027
1152
N conservative
9417
1019
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep1_vs_rep2
rep1_vs_rep2
Rescue Ratio
1.0647764680896252
1.1305201177625124
Self Consistency Ratio
1.1410102944697151
1.2736486486486487
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
31261
27797
25924
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
115.0
122.0
138.0
110.0
110.0
25 percentile
460.0
490.0
550.0
440.0
440.0
50 percentile (median)
460.0
490.0
550.0
440.0
440.0
75 percentile
460.0
490.0
550.0
440.0
440.0
Max size
2815.0
1487.0
1068.0
2787.0
2787.0
Mean
460.65225040785646
490.1038961038961
549.3516046906342
415.1814236111111
436.94195671686447
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
1231191
1359055
871177
Estimated Fragment Length
180
130
130
Cross-correlation at Estimated Fragment Length
0.266385103628973
0.274960539457137
0.250320350532735
Phantom Peak
40
30
30
Cross-correlation at Phantom Peak
0.2608921
0.2709699
0.2445196
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.2483881
0.258251
0.2270267
NSC (Normalized Strand Cross-correlation coeff.)
1.072455
1.064703
1.102603
RSC (Relative Strand Cross-correlation coeff.)
1.439303
1.31376
1.331608
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
0.29153136881048974
0.3020489172023889
0.2703925790299817
Synthetic AUC
0.4885947248175519
0.48896848309777263
0.4872046289711491
X-intercept
0.07246969621389734
0.0661186212400858
0.10354866064747842
Synthetic X-intercept
4.995591286631758e-65
1.3996785574417722e-69
1.7789661043553972e-51
Elbow Point
0.5850271861126353
0.6133147104304884
0.5882755524517385
Synthetic Elbow Point
0.5183551371810522
0.5006506737970494
0.5000565233688058
JS Distance
0.14924686069493662
0.13134774714703928
0.1687143731399045
Synthetic JS Distance
0.26338669848593715
0.24969748534154848
0.27632149190039285
% Genome Enriched
31.995809846859878
35.25355414775278
30.78086496732678
Diff. Enrichment
23.52108451152669
20.964553170850227
25.348069957476444
CHANCE Divergence
0.2007867902950796
0.1788571265710926
0.21646974861157536
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
0.3119437260784111
0.27347456692044575
0.3028082504856588
0.2318816592531292
0.21877849207593628
0.22323942198813643
0.22467363194372608
0.21968443120379566
0.22039818825900417
0.3045051029433532
0.2636866103242995
0.2638981649620754
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
0.1286613212417463
0.12557681130120543
0.1213930114766586
0.10070756180821351
0.0881984006690012
0.09692268935498281
0.1322869922778947
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
0.04222103488485435
0.040242313093643615
0.03724552973497908
0.03002793007137685
0.023279538358198265
0.02344147681823376
0.04521962331722004
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