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
1630738
3184670
2244274
13467896
Distinct Fragments
900900
2090629
1798579
12259949
Positions with Two Read
264684
533636
294288
910990
NRF = Distinct/Total
0.552449
0.656466
0.801408
0.910309
PBC1 = OneRead/Distinct
0.503451
0.634854
0.802324
0.917179
PBC2 = OneRead/TwoRead
1.713587
2.487173
4.903506
12.34324
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
12603
2563
N1
4043
654
N2
10007
2784
N3
6891
1051
Np
16243
3552
N optimal
16243
3552
N conservative
12603
2563
Optimal Set
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2
Conservative Set
rep2_vs_rep3
rep2_vs_rep3
Rescue Ratio
1.2888201221931286
1.3858759266484588
Self Consistency Ratio
2.4751422211229284
4.256880733944954
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
30171
41095
35146
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
89.0
82.0
90.0
82.0
82.0
25 percentile
356.0
330.0
360.0
251.75
330.0
50 percentile (median)
356.0
330.0
360.0
330.0
330.0
75 percentile
356.0
330.0
360.0
330.0
330.0
Max size
13368.0
3824.0
2787.0
10688.0
10688.0
Mean
367.3282622385735
329.0513687796569
359.6791953565128
347.23198198198196
333.7017176629933
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
1630738
3184670
2244274
Estimated Fragment Length
170
155
140
Cross-correlation at Estimated Fragment Length
0.251266432868821
0.438205616521146
0.396866664345704
Phantom Peak
40
40
35
Cross-correlation at Phantom Peak
0.2319788
0.4054939
0.3842881
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.215974
0.3804448
0.3752989
NSC (Normalized Strand Cross-correlation coeff.)
1.16341
1.151824
1.057468
RSC (Relative Strand Cross-correlation coeff.)
2.205114
2.305902
2.399287
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
0.2714607197415334
0.28452811491133684
0.3190977254867476
Synthetic AUC
0.48830136706969757
0.492288839142258
0.4916808319195984
X-intercept
0.09187193101521018
0.04845502814483612
0.04818755239730129
Synthetic X-intercept
9.271886744181683e-62
1.2885554757069688e-143
3.324928087799796e-123
Elbow Point
0.6146592678350433
0.6623058804742704
0.5776298455028145
Synthetic Elbow Point
0.5048398898372308
0.49637957659467635
0.4996446031480059
JS Distance
0.1941605589429008
0.1987916887530947
0.13044598873503713
Synthetic JS Distance
0.2880944811328324
0.2990238132642967
0.24012027210098114
% Genome Enriched
29.98522895125554
26.436983512315862
30.46768334065232
Diff. Enrichment
25.98786248316572
23.752238377623147
17.41484661026324
CHANCE Divergence
0.22215093324179946
0.20439420578521286
0.14885409738712127
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
0.33178150756650837
0.35648585235247776
0.2762345440877898
0.2485216741470027
0.312272736746964
0.23960328814956716
0.24674681415890753
0.31157288584576115
0.24598846193695728
0.3409617429462919
0.32540413607530944
0.3206805088918227
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
0.14646628996815816
0.12995768383861875
0.15751591112017216
0.11145555898313775
0.17886372583061858
0.10139342943242165
0.1825461465302584
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
0.06977637248952206
0.059236848138280805
0.07606921716442719
0.05199883558757207
0.09683195585761542
0.03712174486199075
0.09114431096590503
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