QC Report


general
Report generated at2019-11-27 23:51:23
TitleBY4741--RPB2
Descriptionchipseq of yeast BY4741 for tf RPB2
Pipeline versionv1.3.3
Pipeline typetf
GenomeBY4741
Paired-end per replicate[True, True, True, True]
Alignerbowtie2
Peak callerspp
Control paired-end per replicate[True, True, True, True]

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads58151625050668743148062951023821096602212276819606051150
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads44851933782137693335957628203637548576550872979905604692
Mapped Reads (QC-failed)00000000
% Mapped Reads77.1000000000000174.993.3000000000000191.595.1999999999999995.795.092.60000000000001
Paired Reads58151625050668743148062951023821096602212276819606051150
Paired Reads (QC-failed)00000000
Read129075812525334371574031475511910548301106138409803025575
Read1 (QC-failed)00000000
Read229075812525334371574031475511910548301106138409803025575
Read2 (QC-failed)00000000
Properly Paired Reads34275482889722546339645090703465580459663669404404773966
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads58.957.19999999999999673.571.690.776.390.378.9
With itself44108623716390687949857112263614474571564672502145436122
With itself (QC-failed)00000000
Singletons74331657475386151594230744986247776168570
Singletons (QC-failed)00000000
% Singleton1.31.30.70000000000000010.80.60.80.62.8000000000000003
Diff. Chroms10831038146113615037203395514281
Diff. Chroms (QC-failed)00000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Unpaired Reads00000000
Paired Reads15563981314467248090420529681416051196529128145591975211
Unmapped Reads00000000
Unpaired Duplicate Reads00000000
Paired Duplicate Reads92207105215511556036766905142296671
Paired Optical Duplicate Reads46242073062361054311671986
% Duplicate Reads0.59239999999999990.54050.86870.75790.25960.35130.50550.3377

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads30943562614724491870640748162824750391677256006603937080
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads30943562614724491870640748162824750391677256006603937080
Mapped Reads (QC-failed)00000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads30943562614724491870640748162824750391677256006603937080
Paired Reads (QC-failed)00000000
Read115471781307362245935320374081412375195838628003301968540
Read1 (QC-failed)00000000
Read215471781307362245935320374081412375195838628003301968540
Read2 (QC-failed)00000000
Properly Paired Reads30943562614724491870640748162824750391677256006603937080
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0
With itself30943562614724491870640748162824750391677256006603937080
With itself (QC-failed)00000000
Singletons00000000
Singletons (QC-failed)00000000
% Singleton0.00.00.00.00.00.00.00.0
Diff. Chroms00000000
Diff. Chroms (QC-failed)00000000

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)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Fragments15562781314370248069020528341405680191966027941721964730
Distinct Fragments15471151307291245928620373631402051191323927801261958152
Positions with Two Read90256987208461514034986284130666370
NRF = Distinct/Total0.9941120.9946140.9913720.9924640.9974180.9966550.9949730.996652
PBC1 = OneRead/Distinct0.9941220.994620.9914120.9924880.9974610.996680.9951390.996695
PBC2 = OneRead/TwoRead170.417839186.096751116.960808133.557398399.797313303.451305211.741237306.386342

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
Nt33501746
N138561326
N238491245
N338491632
N439581520
Np35641939
N optimal35641939
N conservative33501746
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep2_vs_rep4rep1_vs_rep3
Rescue Ratio1.06388059701492541.1105383734249714
Self Consistency Ratio1.02831904390750851.310843373493976
Reproducibility Testpasspass

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks4327427142464362

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size202.0211.0185.0192.0203.0202.0
25 percentile810.0844.0740.0770.0706.0677.0
50 percentile (median)810.0844.0740.0770.01376.0810.0
75 percentile810.0844.0740.0770.02207.01494.75
Max size5306.05839.06918.06963.07595.07595.0
Mean965.7492489022418989.4270662608288944.1573245407442943.15933058230171624.98143372872621203.6032547699215

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 Reads2542518222075633258412831811
Estimated Fragment Length8575130115
Cross-correlation at Estimated Fragment Length0.887127562611080.8787450674440290.907019272396070.896813909123608
Phantom Peak55555555
Cross-correlation at Phantom Peak0.88419190.87625180.90212830.8927969
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.79622610.7842160.82808350.8133698
NSC (Normalized Strand Cross-correlation coeff.)1.1141651.120541.0953231.102591
RSC (Relative Strand Cross-correlation coeff.)1.0333721.027091.0660541.050574


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

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 Peaks0.49010488773754540.498161565044723640.47248788604157270.48719377758406760.50728164438739430.52195413359115530.495250378757999040.50467260362185680.51501055470023480.52534263654596050.488313181683630470.50829779798646120.45966047370390630.46424646953696090.4704958306694054

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 Peaks0.434363522864864350.439574981353640650.4359378020298720.43332241463109730.43981908780500210.43656932289944320.46742876385264010.47779306726063630.453552621360170770.46922167773955930.45105703058547053

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 Peaks0.342287576035860850.36030942006047640.34956778398816750.343482194512236660.35115124520135960.35244237720642920.322773785563134960.31759833925110260.342144255013412060.33852915076410810.3762627866822485

For spp raw peaks:


For overlap/IDR peaks: