QC Report


general
Report generated at2022-12-27 19:37:17
Titleelt-3_OP75_L3larva_3_1
DescriptionENCSR962VUP
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}, 'ctl1': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1
Unpaired Reads23531471806498242438712703145944321
Paired Reads00000
Unmapped Reads00000
Unpaired Duplicate Reads2779895590910904742585195349
Paired Duplicate Reads00000
Paired Optical Duplicate Reads00000
% Duplicate Reads11.8135000000000013.09494.49793.35229999999999963.2863

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1
Total Reads20751581750589231534012277295748972
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads20751581750589231534012277295748972
Mapped Reads (QC-failed)00000
% Mapped Reads100.0100.0100.0100.0100.0
Paired Reads00000
Paired Reads (QC-failed)00000
Read100000
Read1 (QC-failed)00000
Read200000
Read2 (QC-failed)00000
Properly Paired Reads00000
Properly Paired Reads (QC-failed)00000
% Properly Paired Reads0.00.00.00.00.0
With itself00000
With itself (QC-failed)00000
Singletons00000
Singletons (QC-failed)00000
% Singleton0.00.00.00.00.0
Diff. Chroms00000
Diff. Chroms (QC-failed)00000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4

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)

rep1rep2rep3rep4ctl1
Total Fragments23406101797993241104212604325896700
Distinct Fragments20670821743792230606212203055728500
Positions with Two Read194378483639178436751145471
NRF = Distinct/Total0.8831380.9698550.9564590.9681640.971476
PBC1 = OneRead/Distinct0.8884270.9707450.9576650.9686320.973011
PBC2 = OneRead/TwoRead9.44783935.00148924.0612232.16312538.316173

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
rep2_vs_rep3
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep4
rep3_vs_rep4
rep3_vs_rep4
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
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt5037380
N1741365
N2672213
N38364307
N44625223
Np4678366
N optimal5037380
N conservative5037380
Optimal Setrep3_vs_rep4rep3_vs_rep4
Conservative Setrep3_vs_rep4rep3_vs_rep4
Rescue Ratio1.07674219752030781.0382513661202186
Self Consistency Ratio1.808432432432432323.615384615384617
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks26160235392573829156

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size166.0624.0116.0110.0111.0111.0
25 percentile536.0624.0464.0440.0424.0424.0
50 percentile (median)536.0624.0464.0440.0424.0424.0
75 percentile536.0624.0464.0440.0424.0424.0
Max size536.0624.0464.0440.0424.0424.0
Mean535.9858562691131624.0463.8879089284327439.93754287282206402.58684210526314422.33154655548935

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

Enrichment / Signal-to-noise ratio


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

rep1rep2rep3rep4
Number of Subsampled Reads2340610179799324110421260432
Estimated Fragment Length130110120120
Cross-correlation at Estimated Fragment Length0.4340404957482070.4135850839357860.4819888588261950.335819420314785
Phantom Peak40303030
Cross-correlation at Phantom Peak0.43213640.41341180.48050840.3343228
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.42600190.40711370.4708070.3243789
NSC (Normalized Strand Cross-correlation coeff.)1.018871.0158961.023751.035269
RSC (Relative Strand Cross-correlation coeff.)1.3104021.027521.1526061.150507


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

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3rep4
AUC0.36254009473611720.35708408863134990.351926630187762550.3240222818586878
Synthetic AUC0.492225051779192960.49153454082787820.49264018085256880.48988681491293
X-intercept0.0440348958790616750.04635955845379010.043270650586253960.05683949651599927
Synthetic X-intercept3.0265429033070455e-1415.972706425184854e-1198.425547697116372e-1585.411145339469899e-83
Elbow Point0.51730227397894430.51203636450709170.5330441306759680.5829655111863162
Synthetic Elbow Point0.5068021997324180.49558665792815960.50166194902979370.49897254783481154
JS Distance0.05310881454754580.0613510378609764960.075390752356386290.11562837160865547
Synthetic JS Distance0.174049683731334110.177785103531205940.193238313735574320.21951078414042935
% Genome Enriched39.33149488773775539.73736301451866437.3652094790361338.07737434849585
Diff. Enrichment11.7043179159557312.6063645309442313.02549177946664617.868061707798443
CHANCE Divergence0.099592884415888390.107373311460349650.110791762258135430.15248473277351868

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep4rep1-pr1rep2-pr1rep3-pr1rep4-pr1rep1-pr2rep2-pr2rep3-pr2rep4-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.199765993721923830.196558986718184560.204120345176086460.24555744793842940.25572896136101440.250638927447317750.249577167932139550.214371237975776420.246535444530006860.248115490338103540.249952922680902160.195748569715767650.097094160038736210.139547210963820780.14001900441509313

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep2_vs_rep3rep2_vs_rep4rep3_vs_rep4rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.032815719648855390.04183019361590790.039554387027712460.040339994919129480.0379656650403538340.0526100800997066560.064141621987337830.062332163631783360.084938713104770790.067198054293740720.048180738940964196

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep2_vs_rep3rep2_vs_rep4rep3_vs_rep4rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0039376746549242110.0041134152352291060.0040325338561853090.0040033568486443410.00401163497636526640.0096723815603483660.00363153070754130550.00144465662699811330.0100672039527671970.0106668491173540730.009634519304051016

For spp raw peaks:


For overlap/IDR peaks: