QC Report


general
Report generated at2019-11-24 01:10:19
TitleAWRI1631--SRB8
Descriptionchipseq of yeast AWRI1631 for tf SRB8
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 Reads653753651472283928384388041459175466601090116525728804978
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads47516824515942266119626857474977004578830996935156321027
Mapped Reads (QC-failed)00000000
% Mapped Reads72.787.767.769.1999999999999984.187.783.271.8
Paired Reads653753651472283928384388041459175466601090116525728804978
Paired Reads (QC-failed)00000000
Read132687682573614196419219402072958773330054558262864402489
Read1 (QC-failed)00000000
Read232687682573614196419219402072958773330054558262864402489
Read2 (QC-failed)00000000
Properly Paired Reads37658983752882210407021426663717114435727672165223826466
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads57.59999999999999472.8999999999999953.655.262.866.061.943.5
With itself46665384462832260445826298464894834569377295276985338574
With itself (QC-failed)00000000
Singletons851445311056738559018217094537165817982453
Singletons (QC-failed)00000000
% Singleton1.31.01.40000000000000011.40000000000000011.40000000000000011.40000000000000011.400000000000000111.200000000000001
Diff. Chroms176024197288262445263351032765
Diff. Chroms (QC-failed)00000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Unpaired Reads00000000
Paired Reads141389913669626819956788191618488190881431347191665993
Unmapped Reads00000000
Unpaired Duplicate Reads00000000
Paired Duplicate Reads769273775426647346667580174165903
Paired Optical Duplicate Reads57858611312465850111711570
% Duplicate Reads0.5440.53970.79560.95359999999999990.28830.397099999999999950.55560.3543

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads28124142719170135313813446923227644380246862346063320180
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads28124142719170135313813446923227644380246862346063320180
Mapped Reads (QC-failed)00000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads28124142719170135313813446923227644380246862346063320180
Paired Reads (QC-failed)00000000
Read1140620713595856765696723461613822190123431173031660090
Read1 (QC-failed)00000000
Read2140620713595856765696723461613822190123431173031660090
Read2 (QC-failed)00000000
Properly Paired Reads28124142719170135313813446923227644380246862346063320180
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0
With itself28124142719170135313813446923227644380246862346063320180
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 Fragments141295813658726799866770681571325185275530445511629062
Distinct Fragments140530613585216745946706351567386184612030296171623881
Positions with Two Read745171425263624538656528144325101
NRF = Distinct/Total0.9945840.9946180.992070.9904990.9974930.9964190.9950950.99682
PBC1 = OneRead/Distinct0.9946280.9946680.9921050.9905480.9975110.9964360.9951570.996835
PBC2 = OneRead/TwoRead187.593075189.201484127.164735106.372458404.523674281.792279208.906943317.337973

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
Nt2641804
N12601736
N22708803
N32657408
N42357422
Np3192785
N optimal3192804
N conservative2641804
Optimal Setpooled-pr1_vs_pooled-pr2rep1_vs_rep2
Conservative Setrep1_vs_rep2rep1_vs_rep2
Rescue Ratio1.208633093525181.024203821656051
Self Consistency Ratio1.14891811624946971.9681372549019607
Reproducibility Testpasspass

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks6268481588986394

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size96.0115.071.072.0102.0102.0
25 percentile384.0460.0284.0290.0318.75410.0
50 percentile (median)384.0460.0284.0290.0410.0410.0
75 percentile384.0460.0284.0290.0410.0410.0
Max size715.01464.0563.0590.01665.01665.0
Mean374.9953733248245444.0336448598131280.0563047875927284.32937128558024379.8320895522388398.2324561403509

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 Reads20750771660806956969918532
Estimated Fragment Length115125130135
Cross-correlation at Estimated Fragment Length0.86500213260510.8401637497244830.7498363456937690.742208214612633
Phantom Peak50505055
Cross-correlation at Phantom Peak0.86851790.84114060.74660530.7361363
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.84559630.82133510.71481580.7043021
NSC (Normalized Strand Cross-correlation coeff.)1.0229491.0229241.0489921.053821
RSC (Relative Strand Cross-correlation coeff.)0.84661710.95067631.1016381.190735


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 Peaks0.335590350496050740.30903327118201510.41911172400745530.34247842628646560.31680092845439650.32544686397182670.417641929142586850.401296356340336670.347180996240949060.342217178195683370.42322722919203980.402669161413914840.26015412519044490.305827871223002370.316338429204142

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 Peaks0.190295931156215990.156633874538332860.145408409395857340.14583857368216010.135066968316334550.13541231489872790.199451076548474020.224126847530680320.185083117908151270.17987687886891570.21583055610034932

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 Peaks0.101283639394007890.082414859672875860.083224880896744290.079217426660027070.078046990952211180.07246190311971180.090779664729303720.105036095573281560.079942326651088070.08564266017794410.10301498988870897

For spp raw peaks:


For overlap/IDR peaks: