ABI公司测序仪

ABI公司测序仪的应用,[e-mail]22006-11-17|©2006AppliedBiosystems提要IDNA测序仪应用概况II基因突变快速扫描：SSCPIII植物AFLP分析IV细菌真菌鉴定：MicroSeqIDV甲基化研究VIBigDyeXTerminatorDNA测序仪应用概况42006-11-17|©2006AppliedBiosystems31031303130xl37303730xlABI遗传分析仪家族使用同样的试剂和软件1416489652006-11-17|©2006AppliedBiosystems测序仪三大功能DNA测序—测定DNA片段的碱基序列片段分析—测定DNA片段的长度和颜色SNP分型—测定多态或点突变62006-11-17|©2006AppliedBiosystems测序仪可以用来做什么？¾DNA测序‰医学测序‰细菌、真菌鉴定‰杂合子、突变检测¾SNP研究‰SNaPshotMultiplex‰SNPlex‰定量PCRTaqMan探针杂交¾片段分析‰基于STR的法医鉴定：HID‰基于STR的基因连锁定位：LMS‰突变筛查：SSCP‰农牧业育种：AFLP72006-11-17|©2006AppliedBiosystems未知序列测序100多种基因组已经完成测序，许多新基因组计划正在进行中许多基因有待发现理解生物学功能解码人类疾病的路径和机制DiagramfromCeleraGenomics82006-11-17|©2006AppliedBiosystems线粒体DNA测序Deleteriousmutationsingene-codingregionsGeneticvariationinpopulationsHumanevolutionDiseaseassociationstudies500-1000copiesofmtDNApercellallowseasierextractionMITOMAP:AHumanMitochondrialGenomeDatabase.://|©2006AppliedBiosystems比较基因组学GeneticconservationandvariationFindinganswerstodiseasemechanismsHowcloselyrelatedaremiceandhumans?Howmanygenesarethesame?–MouseKaryotypeZooWeb-Karyotypeshomepage–HumanKaryotype人类102006-11-17|©2006AppliedBiosystems医学测序VariantSEQr™ResequencingSystemAcompleteapplicationsolutionfordetectingvariantsinthousandsofdisease-relatedhumangenes.SeqScape®Softwarev2.5AllowsautomaticdetectionofheterozygotesMutationProfilingInsertions&DeletionsDevelopmentofdiagnostic/prognosticassaysfordisease112006-11-17|©2006AppliedBiosystems杂合子检测：发现新突变122006-11-17|©2006AppliedBiosystems细菌和真菌鉴定MicroSeq®IDAnalysisSoftwareMicroSeq®50016SBacterialIdentificationKitMicroSeq®D2LSUFungalIdentificationKitDNASequencingofthe16SribosomalRNAgeneClassificationofEubacteria,Yeasts,andFilamentousFungiSequencepolymorphismsBacterialcategoryAbiothreatagentsFoodpathogensSamedayresults132006-11-17|©2006AppliedBiosystemsSAGE™Analysis基因组范围的基因表达图式分析鉴定转录子(mRNA)142006-11-17|©2006AppliedBiosystems基于测序的HLA配型Donor/RecipientcompatibilitypriortotissuetransplantationHLAtypingproducts-distributeddirectlybyCeleraDiagnosticsforresearchpurposesonly.1-866-235-3723:HLA–C6p21.3majorhistocompatibilitycomplex,classI,C.|©2006AppliedBiosystems功能强大的片段分析•片段长度多态性•AFLP和T-RFLP•BAC指纹图谱•相对荧光定量•LOH和QMPSF•构象分析扫描突变•SSCP和CSCE•InVivoScribeB细胞和T细胞clonality•TILLING162006-11-17|©2006AppliedBiosystems相对荧光定量细胞淋巴瘤(celllymphomas)及其他肿瘤中的等位基因丢失染色体不平衡部分和全部染色体丢失非遗传的in-vivoLOHLOH(LossofHeterozygosity)P5317p13.1172006-11-17|©2006AppliedBiosystems寻找新的SNP位点MutationProfiling-SNPDiscoveryandAnalysis182006-11-17|©2006AppliedBiosystemsSNP扫描SNPlex™GenotypingSystemSNaPShot®MultiplexSystemSNPValidationLinkageMappingandAssociationStudiesPharmacogenomicResearchSSCP突变快速扫描202006-11-17|©2006AppliedBiosystemsSSCPmobilitymobilityrfurfuWTWTMUTMUTWildTypeWildTypeMutantMutantATGCATGCAmplifyAmplifyregionofregionofinterestinterestHeatdenatureinHeatdenatureinHiDiHiDiformamideformamideandandNaOHNaOHRapidlychillandRapidlychillandelectrophoresisunderelectrophoresisundernondenaturingnondenaturingconditionsconditionsATGC212006-11-17|©2006AppliedBiosystemsSSCPWorkflowDNAExtractionTissueBloodCellsSampleSetupElectrophoresisSequencesuspectedmutantsBact.DNAAmplificationPostPCRPurificationDataAnalysis222006-11-17|©2006AppliedBiosystemsSSCPdataonABI3130SeriesWTWT+Mut1WT+Mut2Mut1Mut2植物AFLP®分析242006-11-17|©2006AppliedBiosystemsAFLP技术原理252006-11-17|©2006AppliedBiosystemsAFLP工作流程PerformAFLPassayongenomicDNAtemplatesMixAFLPproductwithsizestandardandrunonCEplatformAnalyzedatausingGeneMappersoftwareExport/Print/Reviewfinalresults(Electropherograms,Genotypesdatainbinaryformat262006-11-17|©2006AppliedBiosystemsAB公司AFLP相关产品•Instruments•310•3130•3730•SizeStandard•GS500ROX•Software•GeneMapperv4.0KitsPlantMappingKitSmallGenomes(50-500Mb)RegularGenomes(500-6000Mb)MicrobialFingerprintingKitMicrobialandFungalGenomes272006-11-17|©2006AppliedBiosystems•两个实例•Experiment1-MaizegenomicDNAtemplates，Twoparentalsamples(P1,P2)andfirstgenerationoffspring(F1)fromacrossbetweenP1andP2•Experiment2–Hedysarum(legume)samplesisolatedfromdifferentlocationsinUtah•ABplantmappingkit•Optimalprimersformaize(butincludesinstructionsforotherplantsystems)•Sizestandards–GeneScan™500ROX™•Electrophoresis•Standard36cmPOP-7™PolymerFragmentAnalysisrunmoduleforAppliedBiosystems3130or3730DNAAnalyzers(DyeSetF)•DataAnalysis•NewAFLP®softwareanalysismethodinGeneMapper®Softwarev3.7AFLPDataGeneratedonABSystems282006-11-17|©2006AppliedBiosystemsExpt1:AnalysisofMaizeAFLP®KitSamplesinGeneMapper®SoftwareCommonpeakPolymorphicpeakP1P2F1292006-11-17|©2006AppliedBiosystemsResults:GenotypesFromAFLPAnalysisofMaizeInbredsandaHybrid•Genotypesresultswillbeautomaticallygeneratedinastandardbinaryformat•Tablecanbeprintedorexportedforfurtheranalysis302006-11-17|©2006AppliedBiosystemsExperiment2:AFLPcollaborationwithU.S.customer•Custome