QC Report


general
Report generated at2022-12-26 19:22:43
Titlesea-2_OP193_L3larva_1_1
DescriptionENCSR552OIV
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 Reads53337481453935271855122396275543961
Paired Reads00000
Unmapped Reads00000
Unpaired Duplicate Reads3709803513810164257698240485
Paired Duplicate Reads00000
Paired Optical Duplicate Reads00000
% Duplicate Reads6.95532.41683.73882.57624.3378

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1
Total Reads49627681418797261690921819295303476
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads49627681418797261690921819295303476
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 Fragments53222681452737271417822366215524677
Distinct Fragments49552421417685261338921793365291200
Positions with Two Read285934283308694049496185082
NRF = Distinct/Total0.931040.9758720.9628660.9743880.957739
PBC1 = OneRead/Distinct0.9355470.9785160.9646990.9760440.962032
PBC2 = OneRead/TwoRead16.21304948.96674928.99853942.97573527.502966

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
Nt250665
N1760269
N2547248
N3782216
N482695
Np2777136
N optimal2777136
N conservative250665
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep2_vs_rep3
Rescue Ratio1.10814046288906632.0923076923076924
Self Consistency Ratio1.511147660818713513.8
Reproducibility Testpassfail

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks17599462533062631701

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size125.0129.0476.0524.0149.0149.0
25 percentile490.0400.0476.0524.0410.0410.0
50 percentile (median)490.0400.0476.0524.0410.0410.0
75 percentile490.0400.0476.0524.0410.0410.0
Max size490.0400.0476.0524.0410.0410.0
Mean489.9792601852378399.9895574341124476.0524.0406.5735294117647409.8321930140439

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 Reads5322268145273727141782236621
Estimated Fragment Length130105110130
Cross-correlation at Estimated Fragment Length0.6560927136107210.3717662783189090.5098111954478640.467779394761086
Phantom Peak30353030
Cross-correlation at Phantom Peak0.65586850.36927650.50943560.4675732
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.6509760.35971970.50378540.4617572
NSC (Normalized Strand Cross-correlation coeff.)1.007861.0334891.0119611.013042
RSC (Relative Strand Cross-correlation coeff.)1.0458181.2605231.0664721.035451


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
AUC0.39095888655031990.33183254241315770.375186021247670140.3713798588574375
Synthetic AUC0.49497420112742660.490591041221131340.493079084557011440.49241774328242194
X-intercept0.0361891268994003760.058208702072254540.0409542148479980840.04322900557722815
Synthetic X-intercept0.04.684337571294189e-961.2567621958820313e-1781.3828003481933983e-148
Elbow Point0.45645471869419030.54321403984874640.480359975655748340.48316156002753696
Synthetic Elbow Point0.50365712749518870.49315146902231750.50005531152181070.5076740893748594
JS Distance0.0187596912191641370.103927237902550610.041387288935100090.04644946559410251
Synthetic JS Distance0.15091768388713080.206059184637833880.159711074856433740.16088891587951143
% Genome Enriched39.01325963543485641.4129643117261538.9960989334424242.12992247752646
Diff. Enrichment7.404443547669603617.3600178493548310.1783237513452710.847099939711601
CHANCE Divergence0.06292411888028190.14900576393671850.086523585303887230.0925263167680375

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 Peaks0.121290981162125650.3067429660479970.201905377680309080.219812835339738360.208417560522676040.218111387244695860.244553309055336260.26321834339323440.212665593072253220.20965522879963010.244834743903874330.258752809441924750.034831392034795170.12528042362683430.11741816035204465

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 Peaks0.020779841299101650.0207918265558048320.0189176543993986620.0198442757385400150.0181041774612238950.018140133231333430.055556092890096820.05011076285049940.0558456560774562640.0608383682512125750.022702222808963148

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 Peaks0.00088082692546950230.00115666671407104010.00098762092922768530.00122312227922374540.00070891898977165670.0009062285143031070.00125998233244028340.00133281928281494820.00038709790825741360.000156742038810612080.0023898959634997058

For spp raw peaks:


For overlap/IDR peaks: