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
ctl2
Total Fragments
15240671
23448963
20875245
15281774
16943431
Distinct Fragments
15089766
22626029
20491024
15078127
16747002
Positions with Two Read
130921
634857
327682
177097
170718
NRF = Distinct/Total
0.990099
0.964905
0.981594
0.986674
0.988407
PBC1 = OneRead/Distinct
0.990943
0.968932
0.983339
0.987858
0.989453
PBC2 = OneRead/TwoRead
114.214664
34.532328
61.491388
84.106704
97.06284
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
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
74671
92207
50441
Top 500000 raw peaks from macs2 with p-val threshold 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
idr_opt
overlap_opt
Min size
90.0
115.0
110.0
105.0
105.0
25 percentile
90.0
125.0
110.0
268.0
162.0
50 percentile (median)
92.0
177.0
136.0
386.0
235.0
75 percentile
128.0
297.0
194.0
563.0
375.0
Max size
1109.0
2700.0
1556.0
1911.0
1911.0
Mean
120.07274577814681
260.43937011289813
174.0843758053964
443.91550109072244
303.5650103150549
rep1rep2rep3idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
15000000
15000000
15000000
Estimated Fragment Length
90
115
110
Cross-correlation at Estimated Fragment Length
0.181062779452862
0.234804177557339
0.182362047203375
Phantom Peak
35
40
35
Cross-correlation at Phantom Peak
0.1813714
0.2195042
0.1814161
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.1741223
0.1651067
0.1726048
NSC (Normalized Strand Cross-correlation coeff.)
1.03986
1.422136
1.056529
RSC (Relative Strand Cross-correlation coeff.)
0.9574282
1.281264
1.10735
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
rep1rep2rep3
Jensen-Shannon distance (filtered/deduped BAM)
rep1
rep2
rep3
AUC
0.2497477769123039
0.22359388809402275
0.2639220831031917
Synthetic AUC
0.4850324315824481
0.48799156522528003
0.48723358921681775
X-intercept
0.20086931138861897
0.16703654386743602
0.1530151989539473
Synthetic X-intercept
2.1307284854013305e-37
1.4464424467857623e-58
1.0321627673563e-51
Elbow Point
0.5942007253591808
0.6332177716997957
0.5984653095172403
Synthetic Elbow Point
0.4885104518442233
0.5186775904004263
0.5163263591761047
JS Distance
0.05539363296638151
0.19563146792291408
0.08000764132356082
Synthetic JS Distance
0.2518536296538999
0.34809309883262857
0.26256027951865113
% Genome Enriched
38.17832479616767
26.594970469804025
37.579923465409486
Diff. Enrichment
21.47584658793919
24.031096482016558
18.318015589325938
CHANCE Divergence
0.1903329378077828
0.20535492875209435
0.16058756122877677
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for macs2 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
0.03897717398717116
0.1637390306516292
0.040528460260407746
0.06941180962982181
0.16021323710881094
0.07376138407024113
0.06936735630897234
0.1602281517033232
0.07411711725630422
0.079383907247085
0.09317353204532816
0.0934080475974818
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
0.05301130998182076
0.04854031325146577
0.061726778863737286
0.013678005819735196
0.14132648540721443
0.022930936343189698
0.07582308313510484
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
0.02202807091378155
0.01933056493336402
0.03126486692783061
0.00559376161212275
0.11452376669746071
0.012626894927130742
0.05598194455838486
For macs2 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