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
ctl2
ctl3
ctl4
Total Fragments
1484605
1142674
1647018
798691
1571325
1852755
3044551
1629062
Distinct Fragments
1447349
1121285
1617440
791458
1567386
1846120
3029617
1623881
Positions with Two Read
35572
20624
28495
7094
3865
6528
14432
5101
NRF = Distinct/Total
0.974905
0.981282
0.982041
0.990944
0.997493
0.996419
0.995095
0.99682
PBC1 = OneRead/Distinct
0.974846
0.981271
0.982052
0.99095
0.997511
0.996436
0.995157
0.996835
PBC2 = OneRead/TwoRead
39.664427
53.34969
55.743464
110.557513
404.523674
281.792279
208.906943
317.337973
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
rep4
Number of peaks
14976
9913
13001
12326
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
127.0
85.0
76.0
90.0
87.0
87.0
25 percentile
296.0
340.0
304.0
360.0
248.0
340.0
50 percentile (median)
296.0
340.0
304.0
360.0
274.0
340.0
75 percentile
296.0
340.0
304.0
360.0
297.0
340.0
Max size
365.0
667.0
304.0
360.0
656.0
656.0
Mean
295.9005074786325
336.834157167356
301.3364356587955
355.9570825896479
270.7642487046632
336.1835169432436
rep1rep2rep3rep4idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
2052620
1419949
1971152
1060456
Estimated Fragment Length
110
140
140
65
Cross-correlation at Estimated Fragment Length
0.864584483078785
0.809354455854315
0.858187293036032
0.771144217004943
Phantom Peak
55
55
55
55
Cross-correlation at Phantom Peak
0.8642755
0.8053087
0.856095
0.7713037
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.8576587
0.7829649
0.8463193
0.7526444
NSC (Normalized Strand Cross-correlation coeff.)
1.008075
1.033705
1.014023
1.02458
RSC (Relative Strand Cross-correlation coeff.)
1.046698
1.18107
1.214031
0.9914515
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
rep1rep2rep3rep4
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
0.5540217328334616
0.4468482602772772
0.49676483485578216
0.5224900220723854
0.5697165186638087
0.28295770600672615
0.5350375785893936
0.5079143107670909
0.5765704441189209
0.25610800896822805
0.534042802516344
0.5160831090819299
0.4224876273394088
0.4636246154856868
0.4740497049668914
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
0.2119229605214534
0.2561354556217616
0.2289828009552421
0.21383126152566737
0.1984331601925306
0.23290455730921847
0.32799771305928666
0.12644030877949794
0.2866530241439348
0.2530456616172306
0.30700802638112223
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
0.026305611376378248
0.023688570083168076
0.02369037272909176
0.04274403970147382
0.04318398545384916
0.04223719575593055
0.012167821171719178
0.08213662357565524
0.03853611783883197
0.045252169443352586
0.04497661667795991
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