QC Report


general
Report generated at2019-12-06 07:29:12
TitleENCSR000EGR
DescriptionNFYA ChIP-seq on human K562
Pipeline versionv1.3.5
Pipeline typetf
Genomehg38
Alignerbowtie2
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}, 'ctl2': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3ctl1ctl2
Total Reads556736314983585174918343162373218814423
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads2286682972173673049012128687217523423
Mapped Reads (QC-failed)00000
% Mapped Reads41.09999999999999464.941.867.3000000000000193.10000000000001
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

Marking duplicates (filtered BAM)

rep1rep2rep3ctl1ctl2
Unpaired Reads1820579787480650798981300667911926590
Paired Reads00000
Unmapped Reads00000
Unpaired Duplicate Reads1029271414447784966312669285049
Paired Duplicate Reads00000
Paired Optical Duplicate Reads00000
% Duplicate Reads5.653517.961715.4523999999999992.40392.39

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1ctl2
Total Reads1717652646035942949321269401011641541
Total Reads (QC-failed)00000
Duplicate Reads00000
Duplicate Reads (QC-failed)00000
Mapped Reads1717652646035942949321269401011641541
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

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)

rep1rep2rep3ctl1ctl2
Total Fragments1818466786554750696221279757111751540
Distinct Fragments1716158645630942883461267670611624394
Positions with Two Read87660907975575364112813117699
NRF = Distinct/Total0.9437390.8208340.8458910.9905560.98918
PBC1 = OneRead/Distinct0.9450880.8268250.8436350.9909250.989612
PBC2 = OneRead/TwoRead18.5023965.8792816.28784111.34941997.737797

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
Nt79333872
N11825807
N263763234
N326601559
Np140975210
N optimal140975210
N conservative79333872
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep2_vs_rep3rep2_vs_rep3
Rescue Ratio1.7770074372872811.3455578512396693
Self Consistency Ratio3.49369863013698634.007434944237918
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks1634515638492779

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size70.076.074.079.079.0
25 percentile280.0304.0296.0316.0316.0
50 percentile (median)280.0304.0296.0316.0316.0
75 percentile280.0304.0296.0316.0316.0
Max size481.0692.0318.0767.0767.0
Mean277.7061486693178303.2199905361162295.651591416161285.83435700575814304.829892885011

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 Reads181846678655475069622
Estimated Fragment Length10510090
Cross-correlation at Estimated Fragment Length0.04565936036669330.1065249288272160.0639911888505514
Phantom Peak404035
Cross-correlation at Phantom Peak0.039109660.096757960.05777459
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.022011090.067882730.04957696
NSC (Normalized Strand Cross-correlation coeff.)2.074381.5692491.290745
RSC (Relative Strand Cross-correlation coeff.)1.3830551.3382471.75834


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
AUC0.086012180041008180.178026455438844930.15149868485496332
Synthetic AUC0.453942441848440570.476328747937745930.4709680772830374
X-intercept0.77314131708272850.441209518439877930.5580527264948038
Synthetic X-intercept0.00078191678898914441.2281704150981013e-141.223332523162236e-09
Elbow Point0.77928127861549970.72660676655350420.5824906131164453
Synthetic Elbow Point0.52108796356861680.50238369643129450.4948988238805788
JS Distance0.17997006813484840.12052273400704020.11551036227486353
Synthetic JS Distance0.170109552796248980.240655700976949660.18961652043167432
% Genome Enriched21.90372796231651330.42469899993202341.359068789161995
Diff. Enrichment67.7366424777915933.1370011920664541.84216531505146
CHANCE Divergence0.67410858686754980.291895159122404660.4643960025626234

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 Peaks0.0606286954516980160.118228259451216260.085244190129203440.044752953450407880.078008036703836940.049359570768524390.0465554140186720.077921997511593010.049571448395457720.14252859168842510.103428348591960330.10416660319594206

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 Peaks0.031196005625937680.0272887481326580250.032351627037820980.029539743789778140.0353178515311610360.01664100851887760.03912781450215879

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 Peaks0.025839531215688230.022135513647420660.0281142950785552360.0198614154671609840.0291683480747741730.013508479296063360.031442218568624905

For spp raw peaks:


For overlap/IDR peaks: