QC Report


general
Report generated at2019-12-05 00:33:32
TitleRM11-1A--THP2
Descriptionchipseq of yeast RM11-1A for tf THP2
Pipeline versionv1.3.3
Pipeline typetf
GenomeRM11-1A
Paired-end per replicate[True, True, True, True, True]
Alignerbowtie2
Peak callerspp
Control paired-end per replicate[True, True, True, True, True]

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3rep4rep5ctl1ctl2ctl3ctl4ctl5
Total Reads8236332447314849739484816448594607259791386745678139719781214074810833130
Total Reads (QC-failed)0000000000
Duplicate Reads0000000000
Duplicate Reads (QC-failed)0000000000
Mapped Reads327620437831523089233425868146535545166983627132512144083103055677945377
Mapped Reads (QC-failed)0000000000
% Mapped Reads39.80000000000000484.662.188.478.386.493.086.984.8999999999999973.3
Paired Reads8236332447314849739484816448594607259791386745678139719781214074810833130
Paired Reads (QC-failed)0000000000
Read14118166223657424869742408224297303629895693372839698598960703745416565
Read1 (QC-failed)0000000000
Read24118166223657424869742408224297303629895693372839698598960703745416565
Read2 (QC-failed)0000000000
Properly Paired Reads2523374322857824401963559286358464238697945089238913271876994604827478
Properly Paired Reads (QC-failed)0000000000
% Properly Paired Reads30.59999999999999872.249.173.960.364.775.465.463.444.6
With itself320564837057283039240417311243691845084842619024811938880101333386728650
With itself (QC-failed)0000000000
Singletons7055677424499938556928437082141810772052031722291216727
Singletons (QC-failed)0000000000
% Singleton0.89999999999999991.70000000000000021.01.79999999999999984.81.40000000000000011.21.51.400000000000000111.200000000000001
Diff. Chroms76618596692030108521142798686741032752
Diff. Chroms (QC-failed)0000000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4rep5ctl1ctl2ctl3ctl4ctl5
Unpaired Reads0000000000
Paired Reads90137811494969008721166262153911016663182209745389545133016692074939
Unmapped Reads0000000000
Unpaired Duplicate Reads0000000000
Paired Duplicate Reads549696266077101291029046171426724397182118542
Paired Optical Duplicate Reads2493582402861489672595102412402116
% Duplicate Reads0.60970.83739999999999990.67460.86850.66860.27710.64560.62630000000000010.55160.4117

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4rep5ctl1ctl2ctl3ctl4ctl5
Total Reads1791764227974017895902312266305764033234024390956774210865669164132794
Total Reads (QC-failed)0000000000
Duplicate Reads0000000000
Duplicate Reads (QC-failed)0000000000
Mapped Reads1791764227974017895902312266305764033234024390956774210865669164132794
Mapped Reads (QC-failed)0000000000
% Mapped Reads100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0
Paired Reads1791764227974017895902312266305764033234024390956774210865669164132794
Paired Reads (QC-failed)0000000000
Read189588211398708947951156133152882016617012195478387105432834582066397
Read1 (QC-failed)0000000000
Read289588211398708947951156133152882016617012195478387105432834582066397
Read2 (QC-failed)0000000000
Properly Paired Reads1791764227974017895902312266305764033234024390956774210865669164132794
Properly Paired Reads (QC-failed)0000000000
% Properly Paired Reads100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0
With itself1791764227974017895902312266305764033234024390956774210865669164132794
With itself (QC-failed)0000000000
Singletons0000000000
Singletons (QC-failed)0000000000
% Singleton0.00.00.00.00.00.00.00.00.00.0
Diff. Chroms0000000000
Diff. Chroms (QC-failed)0000000000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
rep5
rep5

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)

rep1rep2rep3rep4rep5ctl1ctl2ctl3ctl4ctl5
Total Fragments89987111476508992421164837153853116229802105576377178232163762033094
Distinct Fragments89440811380638931961154751152825816192282095316375332132015282025906
Positions with Two Read53769385593299001012236551005717329141867012
NRF = Distinct/Total0.9939290.9916460.9932770.9913410.9933230.9976880.9951270.9951050.9953840.996465
PBC1 = OneRead/Distinct0.9939410.9916660.9932960.9913470.9933280.9977130.9951530.9952460.9954720.996497
PBC2 = OneRead/TwoRead165.362537120.253383149.563048115.632222149.976388442.004104207.334295215.562294224.66044287.90773

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
rep1_vs_rep5
rep1_vs_rep5
rep2_vs_rep3
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep4
rep2_vs_rep5
rep2_vs_rep5
rep3_vs_rep4
rep3_vs_rep4
rep3_vs_rep5
rep3_vs_rep5
rep4_vs_rep5
rep4_vs_rep5
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
rep5-pr1_vs_rep5-pr2
rep5-pr1_vs_rep5-pr2
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt2654740
N13547756
N23355746
N33678668
N43787620
N54929474
Np2980851
N optimal2980851
N conservative2654740
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep3_vs_rep4rep2_vs_rep3
Rescue Ratio1.12283345892991711.15
Self Consistency Ratio1.4691505216095381.5949367088607596
Reproducibility Testpasspass

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4rep5
Number of peaks46974784499271816414

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3rep4rep5idr_optoverlap_opt
Min size180.0120.0154.086.0152.0146.0146.0
25 percentile720.0480.0616.0344.0610.0405.0584.0
50 percentile (median)720.0480.0616.0344.0610.0584.0584.0
75 percentile720.0480.0616.0344.0610.01421.0584.0
Max size3514.03445.03398.03222.04544.06451.06451.0
Mean709.5537577176922485.5794314381271608.6398237179487343.6763681938449623.62488306828811231.4900117508814756.8855704697986

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
rep5
rep5
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


Strand cross-correlation measures (trimmed/filtered SE BAM)

rep1rep2rep3rep4rep5
Number of Subsampled Reads14077431368596123492614691902133400
Estimated Fragment Length6514590135120
Cross-correlation at Estimated Fragment Length0.8186556805194870.8141488636565570.7990392182451380.8228553692268370.872696832413182
Phantom Peak5550555050
Cross-correlation at Phantom Peak0.81870890.8117660.79727840.82058270.8721706
Argmin of Cross-correlation15001500150015001500
Minimum of Cross-correlation0.77808110.78022870.76389170.79675540.8504995
NSC (Normalized Strand Cross-correlation coeff.)1.0521471.0434751.0460111.0327581.026099
RSC (Relative Strand Cross-correlation coeff.)0.99868971.0755581.052741.0953831.024282


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
rep5
rep5

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep4rep5rep1-pr1rep2-pr1rep3-pr1rep4-pr1rep5-pr1rep1-pr2rep2-pr2rep3-pr2rep4-pr2rep5-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.36526908677705320.33244887574898890.364044837085589430.37279534448026310.38509405947070290.422402727144869540.39589251405862070.42997174775032520.427978071746008660.450491882628432370.424798131896834630.40059217279163410.426706035132108640.43059010562807710.440470428173362460.25576012821654350.293527453111048660.2914473480357397

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep1_vs_rep5rep2_vs_rep3rep2_vs_rep4rep2_vs_rep5rep3_vs_rep4rep3_vs_rep5rep4_vs_rep5rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2rep5-pr1_vs_rep5-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.21703899919864660.220241652568782840.22382566111655240.200547769566378760.21808716944172380.225766271925919330.200244323746772340.225713382601727350.201552933843825140.20946122339951920.31376230351765080.27674910296788230.30886515905877880.250385552527261130.331530853861147830.23937343068293118

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep1_vs_rep5rep2_vs_rep3rep2_vs_rep4rep2_vs_rep5rep3_vs_rep4rep3_vs_rep5rep4_vs_rep5rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2rep5-pr1_vs_rep5-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.111917371560858330.112595583652390710.103887988602973910.082141038197845250.116326952185913990.113692992609740890.085832606179325090.110452141394354910.083668684890036510.073239960822722830.135153960008126070.12796239922096380.120105722539799620.08977643575609380.074340667966143820.12828189831715786

For spp raw peaks:


For overlap/IDR peaks: