QC Report


general
Report generated at2019-11-26 21:50:27
TitleAWRI1631--NOT3
Descriptionchipseq of yeast AWRI1631 for tf NOT3
Pipeline versionv1.3.3
Pipeline typetf
GenomeAWRI1631
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 Reads581844056347586730144407031659175466601090116525728804978
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads42051054245380598642230897394977004578830996935156321027
Mapped Reads (QC-failed)00000000
% Mapped Reads72.375.388.975.984.187.783.271.8
Paired Reads581844056347586730144407031659175466601090116525728804978
Paired Reads (QC-failed)00000000
Read129092202817379336507220351582958773330054558262864402489
Read1 (QC-failed)00000000
Read229092202817379336507220351582958773330054558262864402489
Read2 (QC-failed)00000000
Properly Paired Reads34380443473726510847424538323717114435727672165223826466
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads59.09999999999999461.675.960.362.866.061.943.5
With itself41493484138096587512630309004894834569377295276985338574
With itself (QC-failed)00000000
Singletons55757107284111296588398217094537165817982453
Singletons (QC-failed)00000000
% Singleton1.01.91.70000000000000021.40000000000000011.40000000000000011.40000000000000011.400000000000000111.200000000000001
Diff. Chroms1131235933068262445263351032765
Diff. Chroms (QC-failed)00000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Unpaired Reads00000000
Paired Reads1487490114909716511548010291618488190881431347191665993
Unmapped Reads00000000
Unpaired Duplicate Reads00000000
Paired Duplicate Reads375182156129753727746667580174165903
Paired Optical Duplicate Reads44038939217665850111711570
% Duplicate Reads2.52221.87629999999999991.8020.90850.28830.397099999999999950.55560.3543

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads28999442255072324280215875043227644380246862346063320180
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads28999442255072324280215875043227644380246862346063320180
Mapped Reads (QC-failed)00000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads28999442255072324280215875043227644380246862346063320180
Paired Reads (QC-failed)00000000
Read11449972112753616214017937521613822190123431173031660090
Read1 (QC-failed)00000000
Read21449972112753616214017937521613822190123431173031660090
Read2 (QC-failed)00000000
Properly Paired Reads28999442255072324280215875043227644380246862346063320180
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0
With itself28999442255072324280215875043227644380246862346063320180
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 Fragments1484605114267416470187986911571325185275530445511629062
Distinct Fragments1447349112128516174407914581567386184612030296171623881
Positions with Two Read355722062428495709438656528144325101
NRF = Distinct/Total0.9749050.9812820.9820410.9909440.9974930.9964190.9950950.99682
PBC1 = OneRead/Distinct0.9748460.9812710.9820520.990950.9975110.9964360.9951570.996835
PBC2 = OneRead/TwoRead39.66442753.3496955.743464110.557513404.523674281.792279208.906943317.337973

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
Nt6039363
N18025141
N21269418
N36481348
N44992366
Np7171386
N optimal7171386
N conservative6039363
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep2_vs_rep4
Rescue Ratio1.18744825302202361.0633608815426998
Self Consistency Ratio6.323877068557922.9645390070921986
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks1497699131300112326

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size127.085.076.090.087.087.0
25 percentile296.0340.0304.0360.0248.0340.0
50 percentile (median)296.0340.0304.0360.0274.0340.0
75 percentile296.0340.0304.0360.0297.0340.0
Max size365.0667.0304.0360.0656.0656.0
Mean295.9005074786325336.834157167356301.3364356587955355.9570825896479270.7642487046632336.1835169432436

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 Reads2052620141994919711521060456
Estimated Fragment Length11014014065
Cross-correlation at Estimated Fragment Length0.8645844830787850.8093544558543150.8581872930360320.771144217004943
Phantom Peak55555555
Cross-correlation at Phantom Peak0.86427550.80530870.8560950.7713037
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.85765870.78296490.84631930.7526444
NSC (Normalized Strand Cross-correlation coeff.)1.0080751.0337051.0140231.02458
RSC (Relative Strand Cross-correlation coeff.)1.0466981.181071.2140310.9914515


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 Peaks0.55402173283346160.44684826027727720.496764834855782160.52249002207238540.56971651866380870.282957706006726150.53503757858939360.50791431076709090.57657044411892090.256108008968228050.5340428025163440.51608310908192990.42248762733940880.46362461548568680.4740497049668914

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 Peaks0.21192296052145340.25613545562176160.22898280095524210.213831261525667370.19843316019253060.232904557309218470.327997713059286660.126440308779497940.28665302414393480.25304566161723060.30700802638112223

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 Peaks0.0263056113763782480.0236885700831680760.023690372729091760.042744039701473820.043183985453849160.042237195755930550.0121678211717191780.082136623575655240.038536117838831970.0452521694433525860.04497661667795991

For spp raw peaks:


For overlap/IDR peaks: