QC Report


general
Report generated at2022-12-26 12:28:55
Titlecey-2_OP358_L4larva_1_1
DescriptionENCSR456YDF
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 Reads184800214547626884577112932
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads22160921121474597573625
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads11.991814.518810.83548.064499999999999

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads162639312435486138606539307
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads162639312435486138606539307
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 Fragments179231314101446642486981057
Distinct Fragments160870412277586014896517772
Positions with Two Read14839313948852405359890
NRF = Distinct/Total0.8975580.8706610.9055190.933637
PBC1 = OneRead/Distinct0.8977090.8709080.9048560.939456
PBC2 = OneRead/TwoRead9.7319157.66563410.38566917.013982

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
Nt8101304
N1813389
N25796225
N3230653
Np7531406
N optimal8101406
N conservative8101304
Optimal Setrep1_vs_rep2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep2rep1_vs_rep2
Rescue Ratio1.07568715973974241.3355263157894737
Self Consistency Ratio3.52688638334778844.245283018867925
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks374322611022262

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size338.0155.0305.0139.0130.0
25 percentile490.0510.0570.0496.0496.0
50 percentile (median)490.0510.0570.0496.0496.0
75 percentile490.0510.0570.0496.0496.0
Max size2821.02976.03706.03886.03886.0
Mean490.907645864501510.83416315587897572.4073308777289642.3029556650247503.28712504629056

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 Reads17923131410144664248
Estimated Fragment Length135135155
Cross-correlation at Estimated Fragment Length0.3798450340850870.3228339850217190.198252965590054
Phantom Peak353535
Cross-correlation at Phantom Peak0.37817710.31932650.1959097
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.37153980.30743060.187103
NSC (Normalized Strand Cross-correlation coeff.)1.0223531.0501031.059592
RSC (Relative Strand Cross-correlation coeff.)1.2512931.2948521.266079


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
AUC0.34073408183916250.31689738223353450.2842632561860546
Synthetic AUC0.491216232176803770.48994452191250290.4856763220954588
X-intercept0.0492501721161809140.057731259291779330.11591351286630813
Synthetic X-intercept2.1641402572870386e-1105.869141220750066e-846.606515627761278e-41
Elbow Point0.55441365754367760.58871117408179760.6331400975823913
Synthetic Elbow Point0.51482588979442620.492361487991432260.5227613979208459
JS Distance0.060795121944273570.098580435743356120.1295122758940286
Synthetic JS Distance0.20418370551652460.23328846239950270.24427808621747937
% Genome Enriched38.291210600361236.5143729483252334.9588417828243
Diff. Enrichment13.4186237620415716.35425110345344323.87255743905397
CHANCE Divergence0.114363515967907450.139520219242859260.20414219306331938

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 Peaks0.27712121240069280.251619559518410250.256457498452415830.282749444476553650.2451549920067420.18104453784250480.242407734420730050.24581600388565620.183996350959502180.190476723555679540.229498117286803330.2214644927952236

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 Peaks0.098772576275166110.081695538866887060.077810127501542130.081685053981417770.088925397330862980.058659955038608150.09351825778797354

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 Peaks0.0210568858554205580.017221132894789340.0204196508353950180.0118187916450697950.0192988127519002070.0210813540546704450.023224059009111026

For spp raw peaks:


For overlap/IDR peaks: