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
1556278
1314370
2480690
2052834
1405680
1919660
2794172
1964730
Distinct Fragments
1547115
1307291
2459286
2037363
1402051
1913239
2780126
1958152
Positions with Two Read
9025
6987
20846
15140
3498
6284
13066
6370
NRF = Distinct/Total
0.994112
0.994614
0.991372
0.992464
0.997418
0.996655
0.994973
0.996652
PBC1 = OneRead/Distinct
0.994122
0.99462
0.991412
0.992488
0.997461
0.99668
0.995139
0.996695
PBC2 = OneRead/TwoRead
170.417839
186.096751
116.960808
133.557398
399.797313
303.451305
211.741237
306.386342
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
4327
4271
4246
4362
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
202.0
211.0
185.0
192.0
203.0
202.0
25 percentile
810.0
844.0
740.0
770.0
706.0
677.0
50 percentile (median)
810.0
844.0
740.0
770.0
1376.0
810.0
75 percentile
810.0
844.0
740.0
770.0
2207.0
1494.75
Max size
5306.0
5839.0
6918.0
6963.0
7595.0
7595.0
Mean
965.7492489022418
989.4270662608288
944.1573245407442
943.1593305823017
1624.9814337287262
1203.6032547699215
rep1rep2rep3rep4idr_optoverlap_opt
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
rep4
Number of Subsampled Reads
2542518
2220756
3325841
2831811
Estimated Fragment Length
85
75
130
115
Cross-correlation at Estimated Fragment Length
0.88712756261108
0.878745067444029
0.90701927239607
0.896813909123608
Phantom Peak
55
55
55
55
Cross-correlation at Phantom Peak
0.8841919
0.8762518
0.9021283
0.8927969
Argmin of Cross-correlation
1500
1500
1500
1500
Minimum of Cross-correlation
0.7962261
0.784216
0.8280835
0.8133698
NSC (Normalized Strand Cross-correlation coeff.)
1.114165
1.12054
1.095323
1.102591
RSC (Relative Strand Cross-correlation coeff.)
1.033372
1.02709
1.066054
1.050574
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
0.4901048877375454
0.49816156504472364
0.4724878860415727
0.4871937775840676
0.5072816443873943
0.5219541335911553
0.49525037875799904
0.5046726036218568
0.5150105547002348
0.5253426365459605
0.48831318168363047
0.5082977979864612
0.4596604737039063
0.4642464695369609
0.4704958306694054
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
0.43436352286486435
0.43957498135364065
0.435937802029872
0.4333224146310973
0.4398190878050021
0.4365693228994432
0.4674287638526401
0.4777930672606363
0.45355262136017077
0.4692216777395593
0.45105703058547053
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
0.34228757603586085
0.3603094200604764
0.3495677839881675
0.34348219451223666
0.3511512452013596
0.3524423772064292
0.32277378556313496
0.3175983392511026
0.34214425501341206
0.3385291507641081
0.3762627866822485
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