QC Report


general
Report generated at2022-12-20 07:43:38
Titleegl-27_OP177_L1larva_2_1
DescriptionENCSR372GNV
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 Reads100247717716084464846686002
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads496445902527618253737
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads4.9521000000000013.33176.1857000000000013.795

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads95283317125834188666432265
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads95283317125834188666432265
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 Fragments99970617685104430406590671
Distinct Fragments95055717098534159146407757
Positions with Two Read435335058424171142740
NRF = Distinct/Total0.9508370.9668330.9387730.972247
PBC1 = OneRead/Distinct0.9515150.968510.9384990.976353
PBC2 = OneRead/TwoRead20.77662932.73782216.14889743.82955

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
Nt6345203
N1353237
N2745985
N31242105
Np6623385
N optimal6623385
N conservative6345203
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep2rep1_vs_rep3
Rescue Ratio1.04381402679275031.896551724137931
Self Consistency Ratio6.0056360708534622.8378378378378377
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks300752799313618

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size624.0223.0155.0120.0120.0
25 percentile624.0630.0430.0480.0480.0
50 percentile (median)624.0630.0430.0480.0480.0
75 percentile624.0630.0430.0480.0480.0
Max size624.0630.0430.0480.0480.0
Mean624.0629.9711356410531429.92443824350124453.53246753246754478.4614223161709

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 Reads9997061768510443040
Estimated Fragment Length105110115
Cross-correlation at Estimated Fragment Length0.2851623441539380.4091461483723850.161920680611905
Phantom Peak303030
Cross-correlation at Phantom Peak0.28387610.40842710.1568671
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.27234650.4004210.1432267
NSC (Normalized Strand Cross-correlation coeff.)1.0470571.021791.13052
RSC (Relative Strand Cross-correlation coeff.)1.1115561.0898131.370483


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
AUC0.325009575325608150.356001265773046970.24411290330190194
Synthetic AUC0.488512793623843870.491437305888908440.48264944382571995
X-intercept0.065548062726946260.046815769522365430.20679741430908583
Synthetic X-intercept4.311188917956582e-643.106909242160553e-1161.1741059825216368e-27
Elbow Point0.52686245560831570.51794820637644150.6752424085232034
Synthetic Elbow Point0.49152635231919960.50224145558622520.5220221409584308
JS Distance0.110600201508082810.070326585381607270.19146154263851656
Synthetic JS Distance0.205791621372675340.178213941606463230.255485241322804
% Genome Enriched37.03802721359881639.6929492039423831.443278400702287
Diff. Enrichment17.63688892901087312.89547222520682730.68840829478092
CHANCE Divergence0.150335934363124060.109835694075383140.26302709023136334

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 Peaks0.266610203466924430.230152348820465920.216785320364985460.195219733972549270.25836163364833490.136115129898344570.19174628895754970.263833206234796340.142012003838936560.173863803634038640.219980390910824030.22276900929876664

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 Peaks0.061660704176855420.0474940358890659150.0483075153309587140.044342502831031250.068060350943574710.052587223598955280.06879007820945036

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 Peaks0.0057037586057306040.0066527639171774820.0062406744908539490.00182508372401039840.0028249725706724870.0121757316182263540.010993806662296119

For spp raw peaks:


For overlap/IDR peaks: