赛默飞世尔精准身份鉴定新一代测序系统介绍课件.pptx

1、The world leader in serving science2016Applied Biosystems Precision ID NGS System for human identification赛赛默飞世尔精准身份鉴定新一代测序系统默飞世尔精准身份鉴定新一代测序系统介绍介绍2Forensic DNA analysis today当前法医当前法医DNADNA分析工作流程分析工作流程CE remains the Gold Standard of HID毛细管电泳毛细管电泳依旧是法医依旧是法医DNADNA分分析的金标准析的金标准Majority of cases can be re

2、solved by STR with CE technology大多数案件可以使用传统的大多数案件可以使用传统的STRSTR和毛细管电泳技术解决和毛细管电泳技术解决Advances in automation allow more efficient uploading of profiles to DNA Databases自动化自动化的优势使得的优势使得DNADNA分型数分型数据上传据上传DNADNA数据库更高效数据库更高效3CE+NGS=Integrated forensics workflow完整法医工作流程完整法医工作流程DNAExtractionDNA提取Quantificatio

3、nBy qPCR定量50-70%ofCaseworkMixtures混合混合Degraded降解降解Investigative leads调查线索调查线索STR by CE传统STR-毛细管检测Converge PlatformConverge平台AnalysisGeneMapper ID-XGMID-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4Complete integrated HID workflow整体解决方案整体解决方案Detection&AnalysisGeneMapper ID-X Expert

4、 System SoftwareGlobalFiler STR Kits3500/3500 xL Genetic Analyzer3130/3130 xl Genetic Analyzer Extraction Quantification Amplification7500 Real-Time PCR InstrumentQuantifiler HP and Trio KitsPrepFiler Forensic DNA Extraction kitProFlex Thermal Cycler AutoMate ExpressDNA Extraction SystemCollectionCo

5、pan NUCLEIC-CARD and 4N6FLOQSwabs Precision ID PanelsNext Gen SequencingY-Filer Plus KitIon Chef System and Ion S5 Systems Converge SoftwareImprove Efficiency.Increase First-Pass Success Rates提高效率，提高首次检出成功率5NGS 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 Sequ

6、encing Sequencing allows greater depth of genetic informationMultiplex hundreds of the same types of markers i.e.SNPsMultiplex different types of markers i.e.SNP+STRSimultaneously analyze multiple samples(barcoding)Target small amplicons to recover DNA with challenging samples6Precision ID NGS Syste

7、m 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 LoadingSimplest 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7Dual Function:-Ion AmpliSeq Library(15 mins)Template Preparation(15 mins)Seq

8、uencing HID NGS System:NGS solution for Forensic CaseworkPipette Sample and primer pools into the Ion Chef System for library prepPipette library into the Ion Chef System for templating,enrichment,chip loadingTransfer Chip to Ion S5 System for SequencingLoad Cartridge into the Ion Chef SystemLoad Ca

9、rtridge into the Ion Chef SystemIon Chef SystemIon S5/S5 XL SystemOnly 3 pipetting steps per sample from DNA to dataOptimized workflow efficiency for targeted sequencing 8Automated Workflows For Walk-Away ConvenienceSingle use reagent cartridges prepackaged and ready to goAutomated system cleaning f

10、ollowing an instrument run,clear the deck and start the system cleaning function to minimize potential contamination Intuitive graphic user interface after loading your library,samples,reagents and consumables on the deck,simply select the run parameters on the touch-screen interface.On board reagen

11、t tracking 2D bar codes associated with all reagents and consumables minimize the potential for user error and ensure every run is set up correctly.Confidence In Sequencing ResultsIon Chef System automation helps to eliminate variation that exists in manual lab workflows to provide consistent result

12、sPrecision ID System for HID Ion Chef System自动化文库模板制备系统9Precision ID System for HIDIon S5 and S5 XL Systems自动化新一代测序系统 Simplest NGS workflow for targeted sequencing 15mins to set up a sequencing run 45mins total hands-on time from DNA to data with Ion Chef System Fastest run time 3.5 hours from seque

13、nce to BAM files.Lowest capital investment Single platform for all targeted applications with flexibility to scale from 5M-80M reads Lowest DNA/RNA input requirements As little as 1 ng using Ion AmpliSeq technology 1ng for HID applications coming soon*Easy setup and training Single day installation

14、and plug and play cartridge-based reagentsSimplest and Fastest targeted sequencing system with the lowest capital investment*The content provided herein may relate to products that have not been officially released,and is subject to change without notice.10Converge NGS Data Analysis新一代测序数据分析软件新一代测序数

15、据分析软件Integrated CE&NGS profile management&data concordanceBackward compatibility for STR allele callingView STR alleles or sequence motif/links SNPs to NCBIAnalysis of single marker sets(STRs)or combination panels(Mixture ID)Data Quality Values similar to GeneMapper ID-XNGS ModuleConverge Case Manag

16、ementSingle repository for case informationData archiving,backup,recovery as well as security and eSigConverge Kinship and PaternitySimple GUI AB algorithm w/configurable analysis settingsAutomated,template based reporting engineThe world leader in serving sciencePrecision ID GlobalFiler STR NGS Pan

17、el精精准身份鉴定准身份鉴定STRSTR试剂盒试剂盒 12 GlobalFiler loci(minus SE33)w/9 addl MPS STRs Expanded CODIS+MPS STR markers Currently available for Ion PGM System,coming in 2017 for Ion S5 System*GlobalFiler STR NGS analysis*The content provided herein may relate to products that have not been officially released,an

18、d is subject to change without notice.1370bp400bpCENGS230bpDesigning smaller amplicons for NGS新一代新一代测序可设计更小的扩增子测序可设计更小的扩增子GlobalFiler multiplexPrecision ID GlobalFiler NGS STR Panel14Example:A 20 allele is not always just a 20Individual A 20 repeats:TCTA GGAA13 GGCA7Individual B 20 repeats:TCTA GGAA

19、14 GGCA6 Precision 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 individuals with the same allele repeat number using sequence dissimilarity within the repeat and flanking

20、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更更有效地分析混合有效地分析混合DNADNAD2S1338 Repeat Structure15 Panel makeup and sample throughput试剂盒组成及通量GlobalFiler STR NGS analysis*The content provided herein may relate to products that have not been officially released,and

21、is subject to change without notice.The world leader in serving sciencePrecision ID mtGenome&mtDNA Control Region 精精准身份鉴定线粒体准身份鉴定线粒体全基因组试剂盒全基因组试剂盒/控制区试剂盒控制区试剂盒 17Bone,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材：骨头、牙齿、头发Precision ID mtDNA Who

22、le 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 11nt18Genome coverage-old bones and teeth陈旧局限检材（骨头、牙齿）基因组覆盖率Performance similar

23、to control mtDNA19New chemistry improvements:Ion Chef library&template prep on Ion S5 system20 Coverage with varying DNA inputsData courtesy of Dr.Bruce Budowle,Institute of Applied Genetics,Department of Molecular and Medical Genetics,University of North Texas Health Science Center Correct SNP call

24、s at all base positions(minimum coverage 10X)Concordant haplotypes obtained from 1ng genomic DNA down to 125pg or less depending on sample.2121mtGenome sequence analysis increases discriminatory potential线粒体基因组测序提高分辨力22Precision ID mtGenome&mtDNA Control Region PanelsThe world leader in serving scie

25、ncePrecision ID Identity and Ancestry Panels精精准身份鉴定个体识别试剂盒和始祖推断试剂盒准身份鉴定个体识别试剂盒和始祖推断试剂盒 24Generate 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 deg

26、raded DNAOnly 1 ng DNA recommended99.99%of concordance for 43 Ken Kidd SNPs25Coverage%Positive Genotype Major AlleleQuality ChecksProfile ComparisonRandom Match Probability(RMP)Y CladesHID SNP Genotyper plug-in for Precision ID Identity Panel26 No suspect and no STR profile matches in DNA database A

27、ncestry or phenotype such 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27Obtain biogeographic ancestry information for investigative leadsPrecision ID Ancestry Panel(A25642)精准始祖推断试剂盒Ready-to-use panel

28、consists of 165 autosomal 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 SNPs28SNP Genotyper plugin for Precision ID Ancestry PanelAdmixture predictionPopulation likelihoodsThe world leader in serv

29、ing scienceApplied Biosystems Precision ID NGS System In development for 2017-GlobalFiler Mixture ID Panel精精准身份鉴定混合物鉴定试剂盒准身份鉴定混合物鉴定试剂盒敬请期待敬请期待30 Non-probative semen stain(epithelial cell fraction)Quantifiler Trio DI=5 some degradation complete STR profile expectedForensic Non-probative sample testin

30、g案件样本实测31Illinois State PoliceResearch and Development LaboratoryForensic Non-probative sample testing案件样本实测32 Non-probative human remains(tissue buried in soil)Quantifiler Trio DI=17 Degradation expectedForensic Non-probative sample testing案件样本实测33STRmhForensic Non-probative sample testing案件样本实测34

31、Evaluation experiments demonstrate that the HID-Ion AmpliSeq Identity Panel is a 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

32、sample.This panel is suitable for personal identification of HAN population from ChinaIdentity SNPs个体识别试剂盒发表文献35The 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:

33、a 97.6%assay conversion rate.Of 3 substitute SNPs added,one failed to give 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 populat

34、ion group differentiations and admixed populations.Analysis of South Asian populations will require extended ancestry-informative SNP panels.EUFORGEN custom Global AIMS Panel欧洲定制始祖试剂盒36 Development of strategies for the complete mtGenome sequencing on the Ion Torrent PGM platform and NGS data analys

35、is 4,15 and 30 samples could be loaded onto Ion 314,Ion 316 and Ion 318 chips,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 Ch

36、inese Han was EMP00670-the first complete mtGenome dataset contributed to EMPOP from East AsiamtDNA sequencing in the Han population37HID NGS Webinars法庭科学新一代测序在线研讨会法庭科学新一代测序在线研讨会Introducing NGS technology for forensics1.Adapting NGS For Forensics Is Not A Paradigm Shift by Dr.Bruce Budowle On demand

37、 recording is available 2.Introducing Mito sequencing on Ion PGM Systemby Dr.Walther Parson On demand recording is available3.Accelerate your forensic DNA analysis with Ion Torrent NGS Automationby Joseph Chang On demand recording is availableAdditional webinars in 2016 ID training delivery:Customer

38、 experience centersCustomer Experience Centers:Targeted NGS Training OfferingsTailor training to meet technical needsEngage HID customers with specialized onsite workshopsDevelop new HID University coursesAccess to NGS data and case examplesView updates to panels and Converge software Initial Planni

39、ng for CECs in ProgressNA Oyster Point,Guilford CTEMEA DarmstadtAPJ Melbourne,DelhiGC Shanghai,BeijingThirteen Centers Globally39For Research,Forensic or Paternity Use Only.When used for purposes other than Human Identification,the instrument and the modules of the cited software are for Research Us

40、e Only.Not for use in diagnostic procedures.2016 Thermo Fisher Scientific Inc.All rights reserved.All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.NUCLEIC CARD and 4N6FLOQSwabs are trademark s of Copan Diagnostics,Inc.Go to us at human_idThank you for your attention!Answers revealed分子刻画，精准打击分子刻画，精准打击3500AnalysisIon S5 and S5 XL Sequencing Systems Maximize results with the proven performance of CE and the power of NGS!