QC Report


general
Report generated at2022-12-26 17:36:50
Titleunc-39_OP186_L3larva_1_1
DescriptionENCSR588ZPD
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 Reads318680539926984756889795129673906
Paired Reads00000
Unmapped Reads00000
Unpaired Duplicate Reads2027492709372352655990635122
Paired Duplicate Reads00000
Paired Optical Duplicate Reads00000
% Duplicate Reads6.36216.78584.94575.71616.565300000000001

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1
Total Reads298405637217614521629235229038784
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads298405637217614521629235229038784
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 Fragments311011639087134552839554969503613
Distinct Fragments296146736991584389219090769014297
Positions with Two Read1258871768811514041305407072
NRF = Distinct/Total0.9522050.9463880.9640620.9514180.948513
PBC1 = OneRead/Distinct0.9545510.9487960.9642920.9519440.951771
PBC2 = OneRead/TwoRead22.45561519.84241427.95561420.95119221.076237

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
Nt697652
N1885332
N2844520
N3128531
N4435342
Np441254
N optimal697654
N conservative697652
Optimal Setrep1_vs_rep2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep2rep3_vs_rep4
Rescue Ratio1.58114233907524931.0384615384615385
Self Consistency Ratio6.8894941634241252.1
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks32626284971517922386

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size494.0295.0656.0233.0490.0490.0
25 percentile616.0616.0656.0796.0490.0490.0
50 percentile (median)616.0616.0656.0796.0490.0490.0
75 percentile616.0616.0656.0796.010607.0490.0
Max size3853.01537.010679.04339.010607.010607.0
Mean617.8282351498805616.0776572972594669.4046379866921798.6240507460024340.925925925926519.8093463302753

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 Reads31101163908713455283955496
Estimated Fragment Length135110150100
Cross-correlation at Estimated Fragment Length0.5371000975102440.5904486549459020.153269218678320.273077208765947
Phantom Peak30303530
Cross-correlation at Phantom Peak0.53705630.59055850.15369680.2733146
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.5321130.58618450.14820780.2630626
NSC (Normalized Strand Cross-correlation coeff.)1.0093721.0072741.0341511.038069
RSC (Relative Strand Cross-correlation coeff.)1.0088650.97487790.92209540.9768445


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
AUC0.373267124129006370.38154255975530230.27369591353772960.3229026792446403
Synthetic AUC0.49351650881587320.49419533642894650.48330655464531370.48832965144624785
X-intercept0.040065538567770750.038502910718669110.16197645481051890.06674197082695543
Synthetic X-intercept9.377510238682775e-2041.3058409724186177e-2546.290012057667916e-304.608001870058969e-62
Elbow Point0.4836822137826570.47096366042081190.61790855334188820.5423735578621335
Synthetic Elbow Point0.50027648993980320.494969139204259560.50681436870801330.5145862773703348
JS Distance0.0433757587129201050.02985631049318010.165777007279370480.11784325645776607
Synthetic JS Distance0.17105764191003710.162885865929947080.23507423125655870.2150772580817568
% Genome Enriched39.5265018090959740.0192384833529237.1272793312032642.60465914556387
Diff. Enrichment9.9849934231506298.2625813101066126.80478769309014518.251039780689826
CHANCE Divergence0.084900305133693430.070240921795585150.230803287644790740.15758235723288583

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 Peaks0.242917693233639040.20818209444400110.2041348012438020.235145454033580140.24707311122847560.235245026414907770.208195292837522830.243299022654576730.244908272498907540.229580628519840070.1395075216404740.211706921979119060.119741122348435020.148532785118409920.15231652539751284

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 Peaks0.054988671040194140.0266730153222773830.035441559680559340.0257460835555177180.034329142568936140.0214821479326674570.072782146179562320.06588144698168420.047509520923916650.064687143349048530.03873810075628278

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 Peaks0.0093525942767315130.0092785981218093030.0094237444256951770.0092461784017597720.0093673192640822530.0095509485181032580.00732291887283616650.0048587214493354090.0294938539726912020.0164284121006321450.009478560975244575

For spp raw peaks:


For overlap/IDR peaks: