2010年微纳尺度分离和分析技术学术会议文章集 邀请报告专题.pdf

2010年微纳尺度分离和分析技术学术会议暨第六届全国微全分析学术会议 AgaroseDroplet Microfluidicsfor HighlyParallelandEfficient EmulsionPCR Chaoyong Yang, Xuefei Leng, andWenhuaZhang College of Chemistry andChemical Engineering, Xiamen University, Xiamen, FJ361005, P.R.China Emulsion PCR(ePCR)hasprovenapromising techniquefor widerangeofapplications suchasraremutationdetectionandnextgenerationDNAsequencing because ofits advantageofmassivelyparallelcolonalamplificationofDNAtemplates. ConventionalePCR methodssufferseveralproblemsincludingpoor amplificationefficiencyandlimitted ampliconlength. Toaddressthese challengingproblems, we have developedanagarose dropletmicrofluidicmethod forhighlyparallelandefficientemulsion PCR.Ourmethod capitalizesonthecapabilityofa microfabricateddropletgeneratortorapidlygenerate monodisperse dropletsandtheuniquethermosensitive sol-gelswitchingpropertyofagarose toavoidtheuse ofmicrobeadsinePCR. Amicrofabricatedagarose dropletgeneratorthat producesuniform, nanoliter-volume agarose emulsion dropletsforgenomicanalysiswas developedandevaluated.Uniformagarose dropletsattherange of50to200μmwere easily obtainedby controllingthemicrofluidic-chipchanneldimension, flowratesofaqueousoroil phase.PCRwasthenefficientlyperformedinagarose solutiondroplets,whichwas subsequentlygelatedtomicrobeadsafterPCRtocaptureampliconstomaintainthe monoclonalityofeachdroplet.Ourresultsindicatedsingle copyDNAcanbeefficiently amplified(90%amplificationefficiency)inagarose droplets.The agarose dropletmethod reportedhereallows uniform, massivelyparallel,highlyefficientmonoclonalamplification andholdsgreatpotentialfor avarietyofapplicationssuchassingle cellexpressionstudy, rare mutantdetection,aswellasnextgenerationhighthroughput sequencing. 2010年微纳尺度分离和分析技术学术会议暨第六届全国微全分析学术会议 基于量子点探针的微纳集成蛋白质芯片技术 樊春海