焦磷酸测序专题知识讲座专家讲座.pptx

pyro=pyrophosphate焦磷酸一个全新基于酶级联反应新型遗传分析技术;一套完整试验方案；很多领域广泛应用……PPiDetectionofthelightCGTTCCACCTAcompletesolutionforaccurategeneticdeterminationPyrosequencing技术流程PeakheightisproportionaltothenumberofincorporatednucleotidesGenotypesareclearlydistinguishedInsertions/deletionsCYP2D6-A2637del(Allele3)LargedeletionMicrosatellites:TG[AATG]nTTTGGGCAAATTri-andtetra-allelicSNPsMultiplepolymorphismsMultiplexingPrincipleofmultiplexingUsercreatesaSNP/mutationsequencedatabaseSoftwarecalculatestheoreticalgenotypingresultsEachDNAfragmentisassignedacolorQuantitativepeakheightsAdvantageandBenefitofPyeosequencing1.AssayDesign2.PCR扩增3.SingleStrandTemplatePreparation真实序列：遗传检测黄金标准快速实时输出：1碱基/分钟96个样本平行处理能力新一代PyroGold试剂：表现大为提升5.PyrogramAnalysisPyrosequencing技术特点WhyPyrosequencingisquantitative？1分子dNTP掺入，释放出1分子PPi，生成1分子ATP，产生单位强度光信号Pyrosequencing：AccurateQuantifyingESGCTAGTAGAGBenefitsofBuilt-inQualityControlIntegratedSoftwarePackage已发表文章使用Pyrosequencing技术发表研究汇报已近千篇发表在Nature、Science上文件快速判定炭疽热细菌Bacillusanthracis炭疽杆菌Bacillusanthracis(B.anthracis)是引发炭疽热严重感染病原细菌。从接触病原体到症状最初出现感染发展过程普通为1到6天。假如确认受到B.anthracis攻击,利用抗生素或解毒剂能够防治其感染,所以快速判定病原微生物种类和其致病力状态含有很主要意义。在起初24-48小时内摄入抗生素能够防治其传染。B.anthracis致病菌株有两个质粒,pXO1含炭疽热毒素产生基因(lef,cya,pag).pXO2编码包装和孢子形成所必须产物.被称为Ba813一段DNA序列是B.anthracis一个特异染色体标识,是B.anthracis区分于与其紧密相关其它土壤杆菌种标志.B.anthracis–assaydesignB.anthracis:rawdataB.anthracis:Result谢谢！