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
2794446
1193074
2067324
3108521
Distinct Fragments
2545779
1139853
1970414
3024930
Positions with Two Read
202016
46668
79237
64412
NRF = Distinct/Total
0.911014
0.955392
0.953123
0.973109
PBC1 = OneRead/Distinct
0.912554
0.956542
0.956753
0.976512
PBC2 = OneRead/TwoRead
11.499886
23.363268
23.791903
45.859157
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
10520
390
N1
7563
99
N2
3768
136
N3
7555
14
Np
7502
323
N optimal
10520
390
N conservative
10520
390
Optimal Set
rep2_vs_rep3
rep1_vs_rep2
Conservative Set
rep2_vs_rep3
rep1_vs_rep2
Rescue Ratio
1.402292721940816
1.2074303405572755
Self Consistency Ratio
2.0071656050955413
9.714285714285714
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
Number of peaks
19890
29204
51304
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
177.0
132.0
280.0
100.0
86.0
25 percentile
456.0
436.0
280.0
340.0
340.0
50 percentile (median)
456.0
436.0
280.0
340.0
340.0
75 percentile
456.0
436.0
280.0
340.0
340.0
Max size
456.0
436.0
280.0
340.0
340.0
Mean
455.97214680744094
435.9697644158334
280.0
337.37692307692305
339.8786121673004
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
2794446
1193074
2067324
Estimated Fragment Length
145
120
70
Cross-correlation at Estimated Fragment Length
0.492279655106282
0.326361537700939
0.441632420079186
Phantom Peak
30
35
40
Cross-correlation at Phantom Peak
0.4902073
0.3246616
0.4375934
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.4839729
0.3100424
0.4276888
NSC (Normalized Strand Cross-correlation coeff.)
1.017164
1.052635
1.032602
RSC (Relative Strand Cross-correlation coeff.)
1.332412
1.116279
1.407797
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
0.3695877691688651
0.32742576382142674
0.3469130887384953
Synthetic AUC
0.49300209337574635
0.48954668901309667
0.4920330381473345
X-intercept
0.04130429579038075
0.057333721635336544
0.04597788274343344
Synthetic X-intercept
1.1010659258393766e-174
1.3151945887200407e-77
1.8437560192231275e-134
Elbow Point
0.5047074968882648
0.5418588229662018
0.5127680879583826
Synthetic Elbow Point
0.502037756826801
0.5127298863921106
0.49956198591305306
JS Distance
0.028771450758231263
0.08771371356479514
0.05926537066667884
Synthetic JS Distance
0.17201849442345968
0.21476668320457687
0.19482536682750795
% Genome Enriched
39.706140490856285
36.14499888296684
40.79923722592793
Diff. Enrichment
10.344771835920769
16.66851648235534
14.03772026754409
CHANCE Divergence
0.08799362039681458
0.14193991532909314
0.12001878589193402
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
0.14640641358037704
0.23411065187693142
0.25575728092496774
0.23778775508292171
0.18752455010306698
0.21279739630320366
0.23117915664447145
0.192359834119516
0.21839336211046698
0.4408202223613352
0.14501781892429866
0.14919706756639434
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
0.04691373559802447
0.05019492029726444
0.0619056038135556
0.06282969801866958
0.053361930220351295
0.04711665662029152
0.046338517734038805
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
0.0057282771337454465
0.0025704663851678503
0.0018836495772069613
0.0028280741693190842
0.01052221762601807
0.0002852091736215901
0.004932673323704776
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