QC Report


general
Report generated at2022-12-27 10:48:58
Titleegl-27_OP177_L2larva_1_1
DescriptionENCSR812VCN
Pipeline versionv1.3.6
Pipeline typetf
GenomeWS245chr
Alignerbwa
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads659543137915317436518559175
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads260973243837453929986924
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads39.56879999999999617.680226.033211.5306

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads398570113531612897227572251
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads398570113531612897227572251
Mapped Reads (QC-failed)0000
% Mapped Reads100.0100.0100.0100.0
Paired Reads0000
Paired Reads (QC-failed)0000
Read10000
Read1 (QC-failed)0000
Read20000
Read2 (QC-failed)0000
Properly Paired Reads0000
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads0.00.00.00.0
With itself0000
With itself (QC-failed)0000
Singletons0000
Singletons (QC-failed)0000
% Singleton0.00.00.00.0
Diff. Chroms0000
Diff. Chroms (QC-failed)0000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3

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)

rep1rep2rep3ctl1
Total Fragments641644133139517000378032660
Distinct Fragments391683111669312739757546913
Positions with Two Read107867160593266363400500
NRF = Distinct/Total0.6104370.8387390.7493810.939528
PBC1 = OneRead/Distinct0.573760.833940.7354810.942488
PBC2 = OneRead/TwoRead2.0834185.7988523.51769617.759983

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
rep2_vs_rep3
rep2_vs_rep3
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
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt108932002
N146129
N25683895
N36049994
Np104021849
N optimal108932002
N conservative108932002
Optimal Setrep2_vs_rep3rep2_vs_rep3
Conservative Setrep2_vs_rep3rep2_vs_rep3
Rescue Ratio1.04720246106517981.0827474310438074
Self Consistency Ratio13.12147505422993534.275862068965516
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks129423106133758

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size94.0104.0100.096.096.0
25 percentile376.0416.0400.0384.0384.0
50 percentile (median)376.0416.0400.0384.0384.0
75 percentile376.0416.0400.0384.0384.0
Max size448.010609.03061.010695.010695.0
Mean375.7630196260238421.8466565789897400.79984003791697463.3256743256743398.51409161847056

rep1
rep1
rep2
rep2
rep3
rep3
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


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

rep1rep2rep3
Number of Subsampled Reads64164413313951700037
Estimated Fragment Length160170155
Cross-correlation at Estimated Fragment Length0.1246444821991960.3137064827822190.323849542327079
Phantom Peak353530
Cross-correlation at Phantom Peak0.11992080.30464010.3138158
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.11189120.28398680.2963655
NSC (Normalized Strand Cross-correlation coeff.)1.113981.1046521.092737
RSC (Relative Strand Cross-correlation coeff.)1.5882831.438981.574993


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

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3
AUC0.255419048198213870.290457746387394860.2989415982644545
Synthetic AUC0.48219736988767070.489469382901596970.49012013421310463
X-intercept0.208976996745180140.065459596130935160.05876436580902529
Synthetic X-intercept3.072381176086196e-261.8641797932843555e-765.202081909153059e-87
Elbow Point0.49949012478472490.61128550417778380.6160130751562054
Synthetic Elbow Point0.48816181677913160.50288835157734980.49072138273388394
JS Distance0.141666678800140280.160700579355077560.1483665869452347
Synthetic JS Distance0.233205547561846640.274238122512876660.26515505305883696
% Genome Enriched32.4234738167602330.15008890387804634.368782465880265
Diff. Enrichment29.62697349040535522.5466591389498921.303898618759227
CHANCE Divergence0.25375641745253030.192651020385998540.18167726059784783

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep1-pr1rep2-pr1rep3-pr1rep1-pr2rep2-pr2rep3-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.17514112953809870.32236047056502330.319771237522504860.109732292947286550.253515320844593050.253142925374615630.1071681260506310.250846460368743140.255075124716799440.26901928312995290.289323447164053950.28906278775240757

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.073205274953180470.074350263917654290.158685979073582450.018240208746267910.13070545997766260.124842407898756470.15355708016126884

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.029841606908607710.0306579383540491470.071418199693441860.0086910705773138970.0547010700104640440.0523539181311941660.06859061172797357

For spp raw peaks:


For overlap/IDR peaks: