QC Report


general
Report generated at2022-12-26 23:07:33
Titlenhr-25_OP33_L2larva_1_1
DescriptionENCSR617FWQ
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 Reads48720973270319419562718623899
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads110165561415010755552638181
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads22.611518.779525.635114.165600000000001

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads37704422656169312007215985718
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads37704422656169312007215985718
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 Fragments46215823159093399408917830145
Distinct Fragments37456512633078309543215961259
Positions with Two Read5975173777435697961390649
NRF = Distinct/Total0.8104690.8334920.7750030.895184
PBC1 = OneRead/Distinct0.8070560.8308760.7690460.900976
PBC2 = OneRead/TwoRead5.0591855.7916654.17786510.341009

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
Nt184553998
N1129912482
N2106731834
N3119442121
Np251665093
N optimal251665093
N conservative184553998
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep1_vs_rep3
Rescue Ratio1.36364128962340821.273886943471736
Self Consistency Ratio1.2171835472688091.3533260632497273
Reproducibility Testpasspass

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks422902908237299

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size110.0109.0111.0110.0110.0
25 percentile440.0436.0444.0419.0440.0
50 percentile (median)440.0436.0444.0440.0440.0
75 percentile440.0436.0444.0440.0440.0
Max size10683.010624.013359.013386.013386.0
Mean441.57786710806334440.2834743140087453.1131933832006480.5383860200275447.9873639036796

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 Reads462158231590933994089
Estimated Fragment Length145175160
Cross-correlation at Estimated Fragment Length0.5827482769312580.5031728493768090.534979509989116
Phantom Peak354030
Cross-correlation at Phantom Peak0.57316490.49267470.5272721
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.55350820.47241780.5057254
NSC (Normalized Strand Cross-correlation coeff.)1.0528271.0651011.057846
RSC (Relative Strand Cross-correlation coeff.)1.4875371.5182521.357709


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
AUC0.3278762779388340.315888778934011570.3233072640124418
Synthetic AUC0.49423493989741870.493129589886742770.49366181874345305
X-intercept0.039513471409454950.043362160218897920.041446803411292414
Synthetic X-intercept3.860584806093472e-2582.7901981681864027e-1812.792481771444423e-213
Elbow Point0.63063572270267030.64097545387365440.6295112744412711
Synthetic Elbow Point0.50901876481027650.49763089256259490.500630156624896
JS Distance0.145600353132905760.163826964224157160.15326302993246338
Synthetic JS Distance0.243294201080139330.25674093223232780.24837895467149593
% Genome Enriched27.4706904471828926.92843875451876627.896902274860693
Diff. Enrichment16.5432991835625217.9751403801345717.307828286325673
CHANCE Divergence0.142866591335758240.155059575463683990.14914187280210617

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 Peaks0.330460725824717640.28678973363517160.31752215974503150.290942016877596850.297928219955801040.297261088846667630.29349397232473010.29657009646980160.29614060188354630.34792052904658090.353785042010398560.34265915634353383

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 Peaks0.18359287723285670.198046169543913850.176632344448852040.169681697795643040.16650973639102030.167127232961290640.23327390256909128

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 Peaks0.088239758249016960.092346524965791780.084490707400675190.069460556613786930.065286132019461110.06828720619267760.10740411093570405

For spp raw peaks:


For overlap/IDR peaks: