QC Report


general
Report generated at2022-12-19 17:44:13
Titlepqm-1_OP201_L1larva_1_1
DescriptionENCSR200QCO
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 Reads2826234120944620765653162971
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads2644205873899069118035
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads9.35594.85664.77083.7318

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads2561814115070819774963044936
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads2561814115070819774963044936
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 Fragments2794446119307420673243108521
Distinct Fragments2545779113985319704143024930
Positions with Two Read202016466687923764412
NRF = Distinct/Total0.9110140.9553920.9531230.973109
PBC1 = OneRead/Distinct0.9125540.9565420.9567530.976512
PBC2 = OneRead/TwoRead11.49988623.36326823.79190345.859157

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
Nt10520390
N1756399
N23768136
N3755514
Np7502323
N optimal10520390
N conservative10520390
Optimal Setrep2_vs_rep3rep1_vs_rep2
Conservative Setrep2_vs_rep3rep1_vs_rep2
Rescue Ratio1.4022927219408161.2074303405572755
Self Consistency Ratio2.00716560509554139.714285714285714
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks198902920451304

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size177.0132.0280.0100.086.0
25 percentile456.0436.0280.0340.0340.0
50 percentile (median)456.0436.0280.0340.0340.0
75 percentile456.0436.0280.0340.0340.0
Max size456.0436.0280.0340.0340.0
Mean455.97214680744094435.9697644158334280.0337.37692307692305339.8786121673004

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 Reads279444611930742067324
Estimated Fragment Length14512070
Cross-correlation at Estimated Fragment Length0.4922796551062820.3263615377009390.441632420079186
Phantom Peak303540
Cross-correlation at Phantom Peak0.49020730.32466160.4375934
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.48397290.31004240.4276888
NSC (Normalized Strand Cross-correlation coeff.)1.0171641.0526351.032602
RSC (Relative Strand Cross-correlation coeff.)1.3324121.1162791.407797


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
AUC0.36958776916886510.327425763821426740.3469130887384953
Synthetic AUC0.493002093375746350.489546689013096670.4920330381473345
X-intercept0.041304295790380750.0573337216353365440.04597788274343344
Synthetic X-intercept1.1010659258393766e-1741.3151945887200407e-771.8437560192231275e-134
Elbow Point0.50470749688826480.54185882296620180.5127680879583826
Synthetic Elbow Point0.5020377568268010.51272988639211060.49956198591305306
JS Distance0.0287714507582312630.087713713564795140.05926537066667884
Synthetic JS Distance0.172018494423459680.214766683204576870.19482536682750795
% Genome Enriched39.70614049085628536.1449988829668440.79923722592793
Diff. Enrichment10.34477183592076916.6685164823553414.03772026754409
CHANCE Divergence0.087993620396814580.141939915329093140.12001878589193402

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 Peaks0.146406413580377040.234110651876931420.255757280924967740.237787755082921710.187524550103066980.212797396303203660.231179156644471450.1923598341195160.218393362110466980.44082022236133520.145017818924298660.14919706756639434

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 Peaks0.046913735598024470.050194920297264440.06190560381355560.062829698018669580.0533619302203512950.047116656620291520.046338517734038805

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 Peaks0.00572827713374544650.00257046638516785030.00188364957720696130.00282807416931908420.010522217626018070.00028520917362159010.004932673323704776

For spp raw peaks:


For overlap/IDR peaks: