QC Report


general
Report generated at2019-11-24 04:12:46
TitleBY4741--STE12
Descriptionchipseq of yeast BY4741 for tf STE12
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 Reads70151026009132546329067406003821096602212276819606051150
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads46717695222560348960056611603637548576550872979905604692
Mapped Reads (QC-failed)00000000
% Mapped Reads66.6000000000000186.963.984.095.1999999999999995.795.092.60000000000001
Paired Reads70151026009132546329067406003821096602212276819606051150
Paired Reads (QC-failed)00000000
Read135075513004566273164533703001910548301106138409803025575
Read1 (QC-failed)00000000
Read235075513004566273164533703001910548301106138409803025575
Read2 (QC-failed)00000000
Properly Paired Reads34258224433526280992046536603465580459663669404404773966
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads48.873.851.469.090.776.390.378.9
With itself45864225099884341655255878063614474571564672502145436122
With itself (QC-failed)00000000
Singletons853471226767304873354230744986247776168570
Singletons (QC-failed)00000000
% Singleton1.22.01.31.09999999999999990.60.80.62.8000000000000003
Diff. Chroms11532960177512815037203395514281
Diff. Chroms (QC-failed)00000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Unpaired Reads00000000
Paired Reads1363029150195394678416118071416051196529128145591975211
Unmapped Reads00000000
Unpaired Duplicate Reads00000000
Paired Duplicate Reads3343330574131853543136766905142296671
Paired Optical Duplicate Reads33434729147261054311671986
% Duplicate Reads2.45280000000000032.03561.39259999999999982.19820.25960.35130.50550.3377

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads26591922942758186719831527522824750391677256006603937080
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads26591922942758186719831527522824750391677256006603937080
Mapped Reads (QC-failed)00000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads26591922942758186719831527522824750391677256006603937080
Paired Reads (QC-failed)00000000
Read11329596147137993359915763761412375195838628003301968540
Read1 (QC-failed)00000000
Read21329596147137993359915763761412375195838628003301968540
Read2 (QC-failed)00000000
Properly Paired Reads26591922942758186719831527522824750391677256006603937080
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0
With itself26591922942758186719831527522824750391677256006603937080
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 Fragments1362152149636094392516065301405680191966027941721964730
Distinct Fragments1328902146595793080915713321402051191323927801261958152
Positions with Two Read3127428893126563344534986284130666370
NRF = Distinct/Total0.975590.9796820.9861050.9780910.9974180.9966550.9949730.996652
PBC1 = OneRead/Distinct0.9757470.9797930.9861580.9781690.9974610.996680.9951390.996695
PBC2 = OneRead/TwoRead41.46166149.71217972.5288445.956914399.797313303.451305211.741237306.386342

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
Nt2150812
N135891111
N22925844
N32285657
N43495471
Np2541636
N optimal2541812
N conservative2150812
Optimal Setpooled-pr1_vs_pooled-pr2rep3_vs_rep4
Conservative Setrep2_vs_rep3rep3_vs_rep4
Rescue Ratio1.18186046511627921.2767295597484276
Self Consistency Ratio1.57067833698030632.358811040339703
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks5639486165525349

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size79.091.075.080.0105.0104.0
25 percentile316.0364.0300.0320.0339.0416.0
50 percentile (median)316.0364.0300.0320.0416.0416.0
75 percentile316.0364.0300.0320.0457.25416.0
Max size1235.0786.0533.0651.03750.03750.0
Mean308.29207306259974359.248508537338296.35027472527474316.1067489250327480.9741379310345430.02558048012594

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 Reads2319319185741613377822006423
Estimated Fragment Length105155135130
Cross-correlation at Estimated Fragment Length0.8732343582114470.8339472420266970.7968920212316230.850035910448012
Phantom Peak55555555
Cross-correlation at Phantom Peak0.86929830.81854010.78907550.8408716
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.83128710.77301340.742690.8084069
NSC (Normalized Strand Cross-correlation coeff.)1.0504611.0788261.0729811.051495
RSC (Relative Strand Cross-correlation coeff.)1.1035491.338421.1685131.282286


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 Peaks0.35087161814566230.34279543204028330.342712984911080650.29486271041934160.40687547194786990.380664410281504440.33047879177377890.34843273432226830.410275752935478130.38129562899540430.335546991317462130.34446730982963450.24657142319170770.271639246598350.2686251117502939

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 Peaks0.175899227068603560.168840697050433530.163299974580818880.211345521987591680.206432088421092260.205962492586072160.280014004253923750.284079424811690240.210836236971119280.240061698478028080.23030484188327888

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 Peaks0.119000555456180160.112922264378312740.111413400615709050.1543156120844670.150586900648659840.151992110639339480.158836593972906060.1982266295767440.14309944633616790.117761879145584560.14688709176324385

For spp raw peaks:


For overlap/IDR peaks: