QC Report


general
Report generated at2019-11-24 14:54:17
TitleBY4741--PHD1
Descriptionchipseq of yeast BY4741 for tf PHD1
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 Reads69457325568928464391865095663821096602212276819606051150
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads35279354805531418951459770003637548576550872979905604692
Mapped Reads (QC-failed)00000000
% Mapped Reads50.886.390.291.895.1999999999999995.795.092.60000000000001
Paired Reads69457325568928464391865095663821096602212276819606051150
Paired Reads (QC-failed)00000000
Read134728662784464232195932547831910548301106138409803025575
Read1 (QC-failed)00000000
Read234728662784464232195932547831910548301106138409803025575
Read2 (QC-failed)00000000
Properly Paired Reads14938743981648403048257945663465580459663669404404773966
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads21.571.586.889.090.776.390.378.9
With itself33773364672722412675259179683614474571564672502145436122
With itself (QC-failed)00000000
Singletons1505991328096276259032230744986247776168570
Singletons (QC-failed)00000000
% Singleton2.19999999999999972.41.40000000000000010.89999999999999990.60.80.62.8000000000000003
Diff. Chroms8123959399518815037203395514281
Diff. Chroms (QC-failed)00000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Unpaired Reads00000000
Paired Reads5228081369926136398920260901416051196529128145591975211
Unmapped Reads00000000
Unpaired Duplicate Reads00000000
Paired Duplicate Reads4595218694139482912936766905142296671
Paired Optical Duplicate Reads8929957174561054311671986
% Duplicate Reads8.78951.36461.02260000000000021.43770.25960.35130.50550.3377

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2ctl3ctl4
Total Reads9537122702464270008239939222824750391677256006603937080
Total Reads (QC-failed)00000000
Duplicate Reads00000000
Duplicate Reads (QC-failed)00000000
Mapped Reads9537122702464270008239939222824750391677256006603937080
Mapped Reads (QC-failed)00000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads9537122702464270008239939222824750391677256006603937080
Paired Reads (QC-failed)00000000
Read14768561351232135004119969611412375195838628003301968540
Read1 (QC-failed)00000000
Read24768561351232135004119969611412375195838628003301968540
Read2 (QC-failed)00000000
Properly Paired Reads9537122702464270008239939222824750391677256006603937080
Properly Paired Reads (QC-failed)00000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0
With itself9537122702464270008239939222824750391677256006603937080
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 Fragments5224521363814135744820151061405680191966027941721964730
Distinct Fragments4771321345246134355119861371402051191323927801261958152
Positions with Two Read3541017443132522740834986284130666370
NRF = Distinct/Total0.9132550.9863850.9897620.9856240.9974180.9966550.9949730.996652
PBC1 = OneRead/Distinct0.9163860.9866420.989910.9858260.9974610.996680.9951390.996695
PBC2 = OneRead/TwoRead12.3478476.092186100.36183271.438485399.797313303.451305211.741237306.386342

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
Nt1984955
N135331288
N22193856
N31626611
N42432747
Np2338970
N optimal2338970
N conservative1984955
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep2_vs_rep4rep2_vs_rep4
Rescue Ratio1.17842741935483871.0157068062827226
Self Consistency Ratio2.17281672816728172.1080196399345335
Reproducibility Testborderlineborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks5164416437773998

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size59.094.099.0104.0119.0119.0
25 percentile236.0376.0396.0416.0409.0476.0
50 percentile (median)236.0376.0396.0416.0476.0476.0
75 percentile236.0376.0396.0416.0550.75476.0
Max size565.0964.0777.0994.02060.02060.0
Mean225.45720371804802370.46685878962535389.42388138734447408.7233616808404520.398969072165485.35543199315657

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 Reads1872651167794513795912044846
Estimated Fragment Length90160175185
Cross-correlation at Estimated Fragment Length0.8230089710147370.8239769203637550.8067054915937760.8515440186479
Phantom Peak55555555
Cross-correlation at Phantom Peak0.80519630.80297090.7776140.8340969
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.71775460.74876520.73949760.8020956
NSC (Normalized Strand Cross-correlation coeff.)1.1466441.1004481.0908831.061649
RSC (Relative Strand Cross-correlation coeff.)1.2037091.3875251.7632281.545202


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 Peaks0.48246325934873420.34102433926964430.309488008141974970.2935623179421130.53908307749089870.35067479159759390.30609418077363520.31989091429881990.53814568758702840.34219586273859710.309628603596930450.304091719413508540.276581663314067970.2919897849159010.29104142770132607

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 Peaks0.147994334398049130.14722806753119270.140410021854692370.244408889507235630.240380843618178630.239467622785304230.4176753569211670.278606856557571170.236150976155538980.245202835708859620.2606700559797028

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 Peaks0.064740419973372450.078712157662958510.038301266258171350.194479709531621660.199523679781414440.186232220115978650.290556268559061860.216962002084024050.17870827626716520.16950606446495450.20714461004542917

For spp raw peaks:


For overlap/IDR peaks: