赛默飞世尔精准身份鉴定新一代测序系统介绍课件

1、2016Applied Biosystems Precision ID NGS System for human identification赛默飞世尔精准身份鉴定新一代测序系统介绍Forensic DNA analysis today当前法医DNA分析工作流程CE remains the Gold Standard of HID毛细管电泳依旧是法医DNA分析的金标准Majority of cases can be resolved by STR with CE technology大多数案件可以使用传统的STR和毛细管电泳技术解决Advances in automation allow mo

2、re efficient uploading of profiles to DNA Databases自动化的优势使得DNA分型数据上传DNA数据库更高效CE + NGS = Integrated forensics workflow完整法医工作流程DNAExtractionDNA提取QuantificationBy qPCR定量50-70% ofCaseworkMixtures混合Degraded降解Investigative leads调查线索STR by CE传统STR-毛细管检测Converge PlatformConverge平台AnalysisGeneMapper ID-XGMID

3、-X分析Precision ID NGS System法医精准身份鉴定新一代测序系统Workflow bottlenecks瓶颈Complete integrated HID workflow整体解决方案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，提高首次检出成功率NGS is a key to getting more information for forensics新一代测序有助于获得更细更多更全的信息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精准身份鉴定新一代测序系统Modular HID Panels(STRs, mtDNA, Ancestry & Identity SNPs)Automated Library Preparation, Templating & Chip LoadingS

7、implest Workflow for Targeted Sequencing Applications试剂盒自动化文库模板制备系统自动化新一代测序系统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自动化文库模板制备系统Precision ID System for HIDIon S5 and S5 XL Systems自动化新一代测序系统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新一代测序数据分析软件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精准身份鉴定STR试剂盒 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新一代测序可设计更小的扩增子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解决方案（NGS STR）不仅会提供每个重复序列的等位基因数目，还有碱基序列Additional resolution for individu

19、als with the same allele repeat number using sequence dissimilarity within the repeat and flanking sequence 同样等位基因数目的重复区和侧翼区序列不同，有助于个体识别率提高Enable more efficient analysis of DNA mixtures更有效地分析混合DNAD2S1338 Repeat StructurePanel makeup and sample throughput试剂盒组成及通量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 精准身份鉴定线粒体全基因组试剂盒/控制区试剂盒 Bone, teeth, and hair are the typical biological sources left from missing persons or remains失踪人员/无名尸体常见生物性检

21、材：骨头、牙齿、头发Precision ID mtDNA Whole Genome Panel, powered by Ion AmpliSeq technology, streamlines the mtGenome sequencing workflowAmpliSeq 技术简化线粒体基因组测序流程Analyze missing person or remains失踪人员/尸源认定Average amplicon length is 163ntAverage amplicon overlap of 11ntGenome coverage - old bones and teeth陈旧局限检

22、材（骨头、牙齿）基因组覆盖率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线粒体基因组测序提高分辨力Precision ID mtGenome & mtDNA Control Region Pa

24、nelsPrecision ID Identity and Ancestry Panels精准身份鉴定个体识别试剂盒和始祖推断试剂盒 Generate identity information beyond STR capabilitiesReady-to-use panel consists of 124 SNPsPi ranges from 1x10-31 6x10-35Precision ID Identity Panel (A25643)精准个体识别试剂盒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分子刻画，更多线索Obtain biogeographic ancestry information for investigative leadsPrecision ID Ancestry Panel (A25642)精准始祖推断试剂盒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精准身份鉴定混合物鉴定试剂盒敬请期待Non-probative semen stain(epithelial cell fraction)Quantifiler Trio DI = 5 some degradation complete STR profile expectedForensic Non-probative sample testing案件样本实测Illinois State PoliceResearch and Development LaboratoryForensic No

29、n-probative sample testing案件样本实测Non-probative human remains (tissue buried in soil)Quantifiler Trio DI = 17Degradation expectedForensic Non-probative sample testing案件样本实测STRmhForensic Non-probative sample testing案件样本实测 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个体识别试剂盒发表文献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欧洲定制始祖试剂盒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法庭科学新一代测序在线研讨会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