賽默飛世爾精準身份鑒定新一代測序系統(tǒng)介紹課件

1、2016Applied Biosystems Precision ID NGS System for human identification賽默飛世爾精準身份鑒定新一代測序系統(tǒng)介紹Forensic DNA analysis today當前法醫(yī)DNA分析工作流程CE remains the Gold Standard of HID毛細管電泳依舊是法醫(yī)DNA分析的金標準Majority of cases can be resolved by STR with CE technology大多數(shù)案件可以使用傳統(tǒng)的STR和毛細管電泳技術(shù)解決Advances in automation allow mo

2、re efficient uploading of profiles to DNA Databases自動化的優(yōu)勢使得DNA分型數(shù)據(jù)上傳DNA數(shù)據(jù)庫更高效CE + NGS = Integrated forensics workflow完整法醫(yī)工作流程DNAExtractionDNA提取QuantificationBy qPCR定量50-70% ofCaseworkMixtures混合Degraded降解Investigative leads調(diào)查線索STR by CE傳統(tǒng)STR-毛細管檢測Converge PlatformConverge平臺AnalysisGeneMapper ID-XGMID

3、-X分析Precision ID NGS System法醫(yī)精準身份鑒定新一代測序系統(tǒng)Workflow bottlenecks瓶頸Complete integrated HID workflow整體解決方案Detection &AnalysisGeneMapper ID-X Expert System SoftwareGlobalFiler STR Kits3500/3500 xL Genetic Analyzer3130/3130 xl Genetic Analyzer Extraction Quantification Amplification7500 Real-Time PCR Inst

4、rumentQuantifiler HP and Trio KitsPrepFiler Forensic DNA Extraction kitProFlex Thermal Cycler AutoMate ExpressDNA Extraction SystemCollectionCopan NUCLEIC-CARD and 4N6FLOQSwabs Precision ID PanelsNext Gen SequencingY-Filer Plus KitIon Chef System and Ion S5 Systems Converge SoftwareImprove Efficienc

5、y. Increase First-Pass Success Rates提高效率，提高首次檢出成功率NGS is a key to getting more information for forensics新一代測序有助于獲得更細更多更全的信息Next-generation Sequencing Sequencing allows greater depth of genetic informationMultiplex hundreds of the same types of markers i.e. SNPsMultiplex different types of markers i.

6、e. SNP + STRSimultaneously analyze multiple samples (barcoding)Target small amplicons to recover DNA with challenging samplesPrecision ID NGS System for human identification精準身份鑒定新一代測序系統(tǒng)Modular HID Panels(STRs, mtDNA, Ancestry & Identity SNPs)Automated Library Preparation, Templating & Chip LoadingS

7、implest Workflow for Targeted Sequencing Applications試劑盒自動化文庫模板制備系統(tǒng)自動化新一代測序系統(tǒng)Dual Function:-Ion AmpliSeq Library (15 mins)Template Preparation (15 mins) Sequencing HID NGS System: NGS solution for Forensic CaseworkPipette Sample and primer pools into the Ion Chef System for library prepPipette libra

8、ry into the Ion Chef System for templating, enrichment, chip loadingTransfer Chip to Ion S5 System for SequencingLoad Cartridge into the Ion Chef SystemLoad Cartridge into the Ion Chef SystemIon Chef SystemIon S5/S5 XL SystemOnly 3 pipetting steps per sample from DNA to dataOptimized workflow effici

9、ency for targeted sequencing Automated Workflows For Walk-Away ConvenienceSingle use reagent cartridges prepackaged and ready to goAutomated system cleaning following an instrument run, clear the deck and start the system cleaning function to minimize potential contamination Intuitive graphic user i

10、nterface after loading your library, samples, reagents and consumables on the deck, simply select the run parameters on the touch-screen interface.On board reagent tracking 2D bar codes associated with all reagents and consumables minimize the potential for user error and ensure every run is set up

11、correctly.Confidence In Sequencing ResultsIon Chef System automation helps to eliminate variation that exists in manual lab workflows to provide consistent resultsPrecision ID System for HID Ion Chef System自動化文庫模板制備系統(tǒng)Precision ID System for HIDIon S5 and S5 XL Systems自動化新一代測序系統(tǒng)Simplest NGS workflow

12、for targeted sequencing15mins to set up a sequencing run 45mins total hands-on time from DNA to data with Ion Chef SystemFastest run time3.5 hours from sequence to BAM files. Lowest capital investment Single platform for all targeted applications with flexibility to scale from 5M - 80M readsLowest D

13、NA/RNA input requirementsAs little as 1 ng using Ion AmpliSeq technology1ng for HID applications coming soon*Easy setup and training Single day installation and plug and play cartridge-based reagentsSimplest and Fastest targeted sequencing system with the lowest capital investment * The content prov

14、ided herein may relate to products that have not been officially released, and is subject to change without notice.Converge NGS Data Analysis新一代測序數(shù)據(jù)分析軟件Integrated CE & NGS profile management & data concordanceBackward compatibility for STR allele callingView STR alleles or sequence motif / links SNP

15、s to NCBIAnalysis of single marker sets (STRs) or combination panels (Mixture ID)Data Quality Values similar to GeneMapper ID-XNGS ModuleConverge Case ManagementSingle repository for case informationData archiving, backup, recovery as well as security and eSigConverge Kinship and PaternitySimple GUI

16、 AB algorithm w/ configurable analysis settingsAutomated, template based reporting enginePrecision ID GlobalFiler STR NGS Panel精準身份鑒定STR試劑盒 GlobalFiler loci (minus SE33) w/ 9 addl MPS STRsExpanded CODIS + MPS STR markersCurrently available for Ion PGM System, coming in 2017 for Ion S5 System*GlobalF

17、iler STR NGS analysis* The content provided herein may relate to products that have not been officially released, and is subject to change without notice.70bp400bpCENGS230bpDesigning smaller amplicons for NGS新一代測序可設(shè)計更小的擴增子GlobalFiler multiplexPrecision ID GlobalFiler NGS STR PanelExample: A 20 allel

18、e is not always just a 20Individual A 20 repeats: TCTA GGAA13 GGCA7Individual B 20 repeats: TCTA GGAA14 GGCA6Precision ID GlobalFiler NGS STR Panel can provide both allele number and base sequence for each repeat structure新一代STR解決方案（NGS STR）不僅會提供每個重復序列的等位基因數(shù)目，還有堿基序列Additional resolution for individu

19、als with the same allele repeat number using sequence dissimilarity within the repeat and flanking sequence 同樣等位基因數(shù)目的重復區(qū)和側(cè)翼區(qū)序列不同，有助于個體識別率提高Enable more efficient analysis of DNA mixtures更有效地分析混合DNAD2S1338 Repeat StructurePanel makeup and sample throughput試劑盒組成及通量GlobalFiler STR NGS analysis* The cont

20、ent provided herein may relate to products that have not been officially released, and is subject to change without notice.Precision ID mtGenome & mtDNA Control Region 精準身份鑒定線粒體全基因組試劑盒/控制區(qū)試劑盒 Bone, teeth, and hair are the typical biological sources left from missing persons or remains失蹤人員/無名尸體常見生物性檢

21、材：骨頭、牙齒、頭發(fā)Precision ID mtDNA Whole Genome Panel, powered by Ion AmpliSeq technology, streamlines the mtGenome sequencing workflowAmpliSeq 技術(shù)簡化線粒體基因組測序流程Analyze missing person or remains失蹤人員/尸源認定Average amplicon length is 163ntAverage amplicon overlap of 11ntGenome coverage - old bones and teeth陳舊局限檢

22、材（骨頭、牙齒）基因組覆蓋率Performance similar to control mtDNANew chemistry improvements: Ion Chef library & template prep on Ion S5 system Coverage with varying DNA inputsData courtesy of Dr. Bruce Budowle, Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas H

23、ealth Science Center Correct SNP calls at all base positions (minimum coverage 10X)Concordant haplotypes obtained from 1ng genomic DNA down to 125pg or less depending on sample.21mtGenome sequence analysis increases discriminatory potential線粒體基因組測序提高分辨力Precision ID mtGenome & mtDNA Control Region Pa

24、nelsPrecision ID Identity and Ancestry Panels精準身份鑒定個體識別試劑盒和始祖推斷試劑盒 Generate identity information beyond STR capabilitiesReady-to-use panel consists of 124 SNPsPi ranges from 1x10-31 6x10-35Precision ID Identity Panel (A25643)精準個體識別試劑盒Small amplicon size (average132;141 nt) for degraded DNAOnly 1 ng

25、DNA recommended99.99% of concordance for 43 Ken Kidd SNPsCoverage% PositiveGenotypeMajor AlleleQuality ChecksProfile ComparisonRandom Match Probability (RMP)Y CladesHID SNP Genotyper plug-in for Precision ID Identity PanelNo suspect and no STR profile matches in DNA database Ancestry or phenotype su

26、ch as eye, hair, or skin color information can provide investigative leads to law enforcement officersHelp generate more investigative leads分子刻畫，更多線索Obtain biogeographic ancestry information for investigative leadsPrecision ID Ancestry Panel (A25642)精準始祖推斷試劑盒Ready-to-use panel consists of 165 autoso

27、mal SNPsSuitable for casework samples or degraded DNA Small amplicon size (average122;130 nt) Only 1 ng DNA recommended99.77% of concordance for 55 Ken Kidd SNPsSNP Genotyper plugin for Precision ID Ancestry PanelAdmixture predictionPopulation likelihoodsApplied Biosystems Precision ID NGS System In

28、 development for 2017- GlobalFiler Mixture ID Panel精準身份鑒定混合物鑒定試劑盒敬請期待Non-probative semen stain(epithelial cell fraction)Quantifiler Trio DI = 5 some degradation complete STR profile expectedForensic Non-probative sample testing案件樣本實測Illinois State PoliceResearch and Development LaboratoryForensic No

29、n-probative sample testing案件樣本實測Non-probative human remains (tissue buried in soil)Quantifiler Trio DI = 17Degradation expectedForensic Non-probative sample testing案件樣本實測STRmhForensic Non-probative sample testing案件樣本實測 Evaluation experiments demonstrate that the HID-Ion AmpliSeq Identity Panel is a

30、well-performed, robust, reliable and high informative NGS-SNP assay. Full profile could be obtained from as low as 100pg input DNA. Compared with STR assays, this panel gives prominence to advantage on detection of degraded sample. This panel is suitable for personal identification of HAN population

31、 from ChinaIdentity SNPs個體識別試劑盒發(fā)表文獻The first custom-built forensic MPS multiplex was built for the EUROFORGEN Global ancestry-informative SNP set analyzed with the Ion PGM System.125/128 SNPs were successfully incorporated: a 97.6% assay conversion rate. Of 3 substitute SNPs added, one failed to giv

32、e usable sequence reads.Five-laboratory evaluations of the assay assessed sequencing performance, forensic sensitivity, and mixture detection.Studies of 14 novel populations indicate good informativeness for 5 continental population group differentiations and admixed populations.Analysis of South As

33、ian populations will require extended ancestry-informative SNP panels.EUFORGEN custom Global AIMS Panel歐洲定制始祖試劑盒Development of strategies for the complete mtGenome sequencing on the Ion Torrent PGM platform and NGS data analysis4, 15 and 30 samples could be loaded onto Ion 314, Ion 316 and Ion 318 c

34、hips, achieving sufficient average coverage of 1500and strand balance of 1.05.Four Perl scripts were developed for primary data analysis and profile comparisons.The EMPOP accession number for the final dataset of 107 Northern Chinese Han was EMP00670- the first complete mtGenome dataset contributed

35、to EMPOP from East AsiamtDNA sequencing in the Han populationHID NGS Webinars法庭科學新一代測序在線研討會Introducing NGS technology for forensicsAdapting NGS For Forensics Is Not A Paradigm Shift by Dr. Bruce BudowleOn demand recording is available Introducing Mito sequencing on Ion PGM Systemby Dr. Walther ParsonOn demand recording is available3.Accelerate your forensic DNA analysis with Ion Torrent NGS Automationby Joseph Chang On demand recording is availableAddi