QC Report


general
Report generated at2022-12-27 01:33:06
Titleegl-27_OP177_L3larva_1_1
DescriptionENCSR669FSH
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 Reads12206213015096418754311627580511
Paired Reads00000
Unmapped Reads00000
Unpaired Duplicate Reads7590512086283929387375284
Paired Duplicate Reads00000
Paired Optical Duplicate Reads00000
% Duplicate Reads6.21859999999999954.0085000000000014.42332.17714.9506000000000006

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1
Total Reads11447162894236134834217757205227
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads11447162894236134834217757205227
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 Fragments11980832882646206624218547383487
Distinct Fragments11309802796466004694142817180147
Positions with Two Read596978062187207151171568
NRF = Distinct/Total0.9439910.9701040.9674650.9820480.97246
PBC1 = OneRead/Distinct0.9441990.9703480.9676890.9822580.974883
PBC2 = OneRead/TwoRead17.88816933.65839731.03990456.90546840.799007

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
Nt4909211
N14315193
N230631
N3152357
N493138
Np6497458
N optimal6497458
N conservative4909211
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep1_vs_rep3
Rescue Ratio1.3234874719902222.170616113744076
Self Consistency Ratio14.1013071895424836.225806451612903
Reproducibility Testborderlinefail

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks2704978471964313587

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size129.0262.0284.0187.0129.0129.0
25 percentile500.0490.0490.0510.0516.0516.0
50 percentile (median)500.0490.0490.0510.0516.0516.0
75 percentile500.0490.0490.0510.0516.0516.0
Max size1123.03037.02767.01358.06347.06347.0
Mean500.11882139820324495.6681534344335491.26497989105536510.21579450945757706.3471615720524529.3546252116362

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 Reads1198083288264620662421854
Estimated Fragment Length120105125120
Cross-correlation at Estimated Fragment Length0.3205044144528190.1074737468794760.2033808595242240.153892904421686
Phantom Peak30303535
Cross-correlation at Phantom Peak0.31932350.1071160.20240230.1535106
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.30579260.10114960.19386620.1447977
NSC (Normalized Strand Cross-correlation coeff.)1.048111.0625221.0490781.062813
RSC (Relative Strand Cross-correlation coeff.)1.0872741.0599681.1146431.043876


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
AUC0.32104691257517320.23288054717230060.286079850514993460.2589958591176447
Synthetic AUC0.48951508491159980.47911327732597380.48566808268633490.4827049163996491
X-intercept0.0578204007678523660.287403868794623430.1133979063651130.18789036153714148
Synthetic X-intercept3.9292802935931675e-776.446502692251736e-197.33625638142448e-417.738008451596911e-28
Elbow Point0.55727364518356230.6118893398624730.63253036089859480.6555759092305912
Synthetic Elbow Point0.5157432968680690.49628826303854150.48657365479737790.5012964203784969
JS Distance0.128666410056333140.17918324396671730.16569583243501190.1745683559016435
Synthetic JS Distance0.225565105666885060.231254703296359420.243743650429532640.24022644659552497
% Genome Enriched34.80658022341151538.204247133524134.6517326564738733.24533362067933
Diff. Enrichment18.00988763500638334.2966129627654323.85943917801999328.71025497508713
CHANCE Divergence0.153380338931049160.301879047946185860.203872387760679730.24600815107132804

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 Peaks0.25009085222884980.145213061850647670.222883111675466150.18769249007172070.199132361214484630.094449665542594940.164075998722053060.108403512765069610.218328388875494020.104193876070236540.134774940422049880.14083845851095610.189681529539397680.232322021543694830.23674796326548134

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 Peaks0.052503505916626610.086238867221430980.075652477102709680.046149323093856520.042520906925860850.063334085203796710.06932549208712030.036842268928177790.0491912571334495040.0355734692667891640.10195646953487025

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 Peaks0.0197700086296371150.0246760646425017920.024389354972084280.0199963796829752370.019313621908506410.0220474877065129670.0167185572666058640.032699543574629520.021863686524320970.013037757097978780.03214144991672056

For spp raw peaks:


For overlap/IDR peaks: