QC Report


general
Report generated at2022-12-26 11:18:00
Titlenhr-76_OP203_L3larva_1_1
DescriptionENCSR415EON
Pipeline versionv1.3.6
Pipeline typetf
GenomeWS245chr
Alignerbwa
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'rep4': {'paired_end': False}, 'rep5': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


Marking duplicates (filtered BAM)

rep1rep2rep3rep4rep5ctl1
Unpaired Reads329082439946381587493945125111198116063861
Paired Reads000000
Unmapped Reads000000
Unpaired Duplicate Reads283040315864232787336907796331375437
Paired Duplicate Reads000000
Paired Optical Duplicate Reads000000
% Duplicate Reads8.60097.907200000000000514.66379999999999835.64687.16139999999999958.5623

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4rep5ctl1
Total Reads300778436787741354706608218103234814688424
Total Reads (QC-failed)000000
Duplicate Reads000000
Duplicate Reads (QC-failed)000000
Mapped Reads300778436787741354706608218103234814688424
Mapped Reads (QC-failed)000000
% Mapped Reads100.0100.0100.0100.0100.0100.0
Paired Reads000000
Paired Reads (QC-failed)000000
Read1000000
Read1 (QC-failed)000000
Read2000000
Read2 (QC-failed)000000
Properly Paired Reads000000
Properly Paired Reads (QC-failed)000000
% Properly Paired Reads0.00.00.00.00.00.0
With itself000000
With itself (QC-failed)000000
Singletons000000
Singletons (QC-failed)000000
% Singleton0.00.00.00.00.00.0
Diff. Chroms000000
Diff. Chroms (QC-failed)000000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
rep5
rep5

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)

rep1rep2rep3rep4rep5ctl1
Total Fragments320867839138421564984930622109454615759633
Distinct Fragments298485836561421342295601079102073014663381
Positions with Two Read18427321327817059315224463695875459
NRF = Distinct/Total0.9302450.9341570.8577050.645890.932560.930439
PBC1 = OneRead/Distinct0.9329620.9369250.8553280.6220610.9331370.936159
PBC2 = OneRead/TwoRead15.11214316.0613476.7300652.45597914.9537815.680075

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


Replication quality metrics


IDR (Irreproducible Discovery Rate) plots

rep1_vs_rep2
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep3
rep1_vs_rep4
rep1_vs_rep4
rep1_vs_rep5
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep4
rep2_vs_rep5
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep4
rep3_vs_rep5
rep3_vs_rep5
rep4_vs_rep5
rep4_vs_rep5
rep1-pr1_vs_rep1-pr2
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep3-pr1_vs_rep3-pr2
rep4-pr1_vs_rep4-pr2
rep4-pr1_vs_rep4-pr2
rep5-pr1_vs_rep5-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt2219213
N1533080
N2635666
N34966268
N499959
N5506533
Np2622139
N optimal2622213
N conservative2219213
Optimal Setpooled-pr1_vs_pooled-pr2rep3_vs_rep4
Conservative Setrep1_vs_rep3rep3_vs_rep4
Rescue Ratio1.1816133393420461.5323741007194245
Self Consistency Ratio6.3623623623623628.121212121212121
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4rep5
Number of peaks1801819755301741982725209

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4rep5idr_optoverlap_opt
Min size198.0151.0120.099.0800.0128.0126.0
25 percentile684.0604.0470.0390.0800.0504.0504.0
50 percentile (median)684.0604.0470.0390.0800.0504.0504.0
75 percentile684.0604.0470.0390.0800.0504.0504.0
Max size10679.010661.02733.0965.010695.010648.010648.0
Mean692.9543234543235612.6592761326247470.49423344601314390.12170272860243808.48625490896111399.9483568075118573.8318077803203

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
rep5
rep5
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


Strand cross-correlation measures (trimmed/filtered SE BAM)

rep1rep2rep3rep4rep5
Number of Subsampled Reads3208678391384215649849306221094546
Estimated Fragment Length150105150135150
Cross-correlation at Estimated Fragment Length0.5343035425681790.5847016891897960.3396030193109210.1809509249911580.292868784924775
Phantom Peak3530403030
Cross-correlation at Phantom Peak0.53448210.58524920.33533810.17542990.2938288
Argmin of Cross-correlation15001500150015001500
Minimum of Cross-correlation0.52981560.58104220.32352090.16498160.2826909
NSC (Normalized Strand Cross-correlation coeff.)1.0084711.0062981.049711.0967951.036004
RSC (Relative Strand Cross-correlation coeff.)0.96173020.8698611.3609111.5284150.9138073


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.


rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
rep5
rep5

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3rep4rep5
AUC0.37389070353051890.38454733026826870.327057692060665650.287886144582595370.33896959497934365
Synthetic AUC0.4935415101957270.494162036019706760.490367070598956230.48560457695726650.4889628272903833
X-intercept0.039949339171847280.03805355344957090.053869080913493040.117131190769181590.058335863711185335
Synthetic X-intercept2.3903039552575013e-2051.1026321797741657e-2511.371971671175897e-911.707136435521003e-401.657908682326754e-69
Elbow Point0.489967717705719730.47204065651319260.55079467182331030.62635441803972960.5533416968980231
Synthetic Elbow Point0.50616919187217520.50731987583208640.50202561475051440.52121846612950130.50596580614627
JS Distance0.061146033131389060.043017883861597940.123361457598634360.15828008193878850.10074520244374248
Synthetic JS Distance0.171570493322034780.15961715103899450.21932650611162880.233175493140254320.19463691833412292
% Genome Enriched37.75869784092180538.2950713566554335.9078596199638835.7102903808351840.72523292794017
Diff. Enrichment8.7855067209270677.65017018662068616.3967425633848124.14351451581729416.29840187268098
CHANCE Divergence0.074666567457128810.06501319347763880.139609666872401280.206563967578905130.13940253179198764

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep4rep5rep1-pr1rep2-pr1rep3-pr1rep4-pr1rep5-pr1rep1-pr2rep2-pr2rep3-pr2rep4-pr2rep5-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.16167849819002960.15186418083850760.26571743241707060.211943743854999370.241133803717351130.2222526617602860.21418983607038650.221122516619842250.121591271550661110.247577367321872080.221977376034981240.224950486221768440.224783827634925960.13414269225836790.241699504430676480.070697068632686170.116669885755069040.1138941708334065

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep1_vs_rep5rep2_vs_rep3rep2_vs_rep4rep2_vs_rep5rep3_vs_rep4rep3_vs_rep5rep4_vs_rep5rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2rep5-pr1_vs_rep5-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0254124478533500380.0291929315015859620.023020338097239880.0210492231324036880.0266748125096185340.020754960580799290.0214607155878589060.0293645932638767660.0242057544906283220.0195701639049642470.0566975554095639840.05556878460052180.076096953877815550.028047509281211670.058996578673083110.03250552839700759

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep1_vs_rep5rep2_vs_rep3rep2_vs_rep4rep2_vs_rep5rep3_vs_rep4rep3_vs_rep5rep4_vs_rep5rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2rep5-pr1_vs_rep5-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0093299510526419080.008698872010766560.0088601018609085270.0082801495171883830.0084347690467607880.0084613136152979340.0081888444643213110.0113393852195297780.0085628440077960470.0085317548438673270.0106231032547549950.0075612146872844050.0170708626078278250.0101641187863562070.0116530472282602390.010459076434930173

For spp raw peaks:


For overlap/IDR peaks: