QC Report


general
Report generated at2021-03-26 16:27:02
Titlemyc_hepg2
Descriptionchipseq of myc_hepg2
Pipeline versionv1.3.6
Pipeline typetf
Genomehg38
Alignerbowtie2
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callermacs2

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3ctl1
Total Reads726748875853921195004512326775
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads683324571445861056397011595209
Mapped Reads (QC-failed)0000
% Mapped Reads94.094.1999999999999988.494.1
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

Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads5315539552169280743239292850
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads1517401742455267732703083
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads2.85463.15566.52409999999999929.0878

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads5315539552169280743239292850
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads5315539552169280743239292850
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 Fragments5300305551357880460499262390
Distinct Fragments5153768534111175333736576327
Positions with Two Read1371261611844570781563444
NRF = Distinct/Total0.9723530.968720.9362820.710003
PBC1 = OneRead/Distinct0.9725290.9688250.9357650.686559
PBC2 = OneRead/TwoRead36.55169732.10368915.4228952.887878

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
Nt371945419
N19457625520
N2466174771
N38866911609
Np297437771
N optimal371947771
N conservative371945419
Optimal Setrep1_vs_rep3pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep1_vs_rep3
Rescue Ratio1.25051272568335411.4340284185274035
Self Consistency Ratio2.0287877812815075.348983441626493
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks499601226728472185

Top 500000 raw peaks from macs2 with p-val threshold 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size70.080.080.077.077.0
25 percentile70.080.080.0193.0104.0
50 percentile (median)75.080.080.0273.0150.0
75 percentile115.086.091.0400.0227.0
Max size1641.01354.01576.02277.02277.0
Mean102.4987520040992798.50551321407148100.97283056429154319.70801698623086190.3113136527397

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 Reads530030555135788046049
Estimated Fragment Length708080
Cross-correlation at Estimated Fragment Length0.08752210805071750.08022212025161290.109520146886115
Phantom Peak403540
Cross-correlation at Phantom Peak0.085625970.078953050.101537
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.069539870.070705480.08787084
NSC (Normalized Strand Cross-correlation coeff.)1.2585891.1345961.246376
RSC (Relative Strand Cross-correlation coeff.)1.1178741.1538731.58415


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
AUC0.148165154196257230.164273378182251320.1678126717292743
Synthetic AUC0.474215709498406470.47471135981356210.47913649884900006
X-intercept0.52556951092237380.49472375171443080.42938144536014045
Synthetic X-intercept2.762667646887698e-128.774203266191945e-135.83550938126397e-19
Elbow Point0.5580967118916520.52905435395448710.6849505964019945
Synthetic Elbow Point0.54468807818814350.53079525710348230.49028859969785354
JS Distance0.127222027109829580.103537235708873130.11187965202618459
Synthetic JS Distance0.24710907693182120.218516265728740280.2745718592060635
% Genome Enriched33.37771859742565542.09842332345661530.842360336368326
Diff. Enrichment44.47793436357835640.49925993770301639.75161149983176
CHANCE Divergence0.402950949715589150.40153552878525080.3527082182396602

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for macs2 raw peaks

rep1rep2rep3rep1-pr1rep2-pr1rep3-pr1rep1-pr2rep2-pr2rep3-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.273201833341830460.148789718803584130.24553464110861060.33766428246236510.302969814325029340.305192360375927450.337691123645433470.30251596793156880.305445831860557460.072830979410787710.082766325520755660.08274656675454241

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 Peaks0.056779363557325850.064146658703985930.0581673510278425540.108229475881937850.053848530486669670.09831350566480930.05982728865116003

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 Peaks0.0216956258591969770.0254613661045517450.0221565610102691730.068335873370508620.018232817042312390.04105681677584610.031619876399369404

For macs2 raw peaks:


For overlap/IDR peaks: