具有单一热源的对流双温PCR微流控装置的研究-章春笋

2011304TransducerandMicrosystemTechnologiesPCR*510631PCRPCRPCRPCRPCR。PCRPCR。PCR45min112bpDNA。DNATK323O351Q819A1000—9787201104—0069—03Studyofaconvectivetwo-temperaturePCRmicrofluidicsdevicewithasingleheatsource*ZHANGChun-sunMOEKeyLaboratoryofLaserLifeScience＆InstituteofLaserLifeScienceSouthChinaNormalUniversityGuangzhou510631ChinaAbstractToovercomeshortcomingsoftheconventionalcontinuous-flowPCRmicrosystemsusuallyneedmultipleheatsourcestoprovidetemperaturesforPCRandtheintegratedornonintegratedpumpingsystemsarealsorequiredtodrivethePCRsolutiontoflowcontinuouslyandthusthePCRisrealized．Aconvectivetwo-temperaturePCRmicrofluidicswithasingleheatsourceisstudied．ThetemperaturesforthisPCRmicrosystemareformedonthegroovedcopperflakebythecombinedfintechnology．AndthelargetemperaturedifferencesneededforPCRinducethedifferencesintheflowdensitywhichundercertainconditionscanbeusedtotransportthePCRsolutionwithinaclosed-loopchannel．TheexperimentdemonstratesthatthedevelopedmicrofluidicPCRcanamplifythe112bpEscherichiacoliDNAfragmentin45min．KeywordsmicrofluidicsconvectivepolymerasechainreactionPCRDNAamplificationsingleheatsource0PCRDNA、。PCR20～401～2h。PCR。PCR1。PCR2PCRPCR12。PCR。PCRDNA。DNAPCR500bp700bpλ-DNA1．7min3．2min3。PCR、、。PCR。2010—10—13*3070015561072030“”IRT0829—U0931005“863”2007AA10Z2049630PCRPCRDNA。—Ray-leigh-Bénard。PCR2PCR45PCR5～9。、PCRPCR。PCRDNA。PCR。PCR2578369PCR2～3。AgrawalN95℃8。PCR72℃55℃。3。。、PCRDNA。11．1TaqDNA5U/μL10PCR500mmol/LKCl100mmol/LTris-HClpH8．80．8%NP—4025mmol/LMgCl2dNTPsdATPdGTPdCTPdTTP10mmol/LPCR/5’—GGAATCAGGCGTCTGGGTCA—3’and5’—GCCGTTAGTCGCTTCGTCATA—3’ddH2ODNADNAmarkerBSAV＞98%GoldViewTME．coliK12DNATRIzolBiophotometerGelDocXRPT-FE0．5mm0．9mm。1．2PCRPCR1DNA2DNA3DNA。dNTPsDNA300bpTaq。PCR。PCR1。90～97℃DNA50～70℃/ββ=60°PCRPCR。PCR2LabVIEW、、72mm×40mm×4mm、、8mm40mm100W、K0．12mmOmega、、。181．1mm21．1mm40mm×45mm。716mm1．6mm24．8mm。1～3PCR。1LabVIEW8．0//PID。PCR。“0”PTFE180mm35μLPCR。1PCRFig1Schematicdiagramofconvectivetwo-temperaturePCRinaclosed-loopchannel1．3PCRPCR1PCR1．5mmol/LMgCl20．2mmol/LdNTPs0．5μmol/L/0．25μg/μLBSA0．1U/μLTaqDNA4ng/μLDNA12090604530min。PCR0．2mL074PCR2PCRFig2Schematicdiagramoftheconvectivetwo-temperaturePCRmicrofluidicsystemwithasingleheatsourcePCR4℃。PCR1．2%W/VGoldViewTM20mL1．0μLGoldViewTM125V35minPCR。22．1PCRPCRPCR。3a3b。197℃。P=0．00W1786．7±0．2℃11812℃PCR。118。P=0．08W1773．1±0．1℃P=0．72W1756．3±0．2℃。PCR。。PCR。2．2PCRDNAPCR1PCRDNA。4PCR。PTFE2180．32W21891．6±3PCRFig3Temperaturesforconvectivetwo－temperaturePCRformedonthegroovedcopperflakewithsingleheatsource0．2℃60．5±0．2℃。1PCRDNA2120min30min。60min45min12090min60minPCR60min3PCR30minDNA。PTFEPCR30minDNA。PCR10～12PCR。4PCRFig4Effectofdifferentreactiontimesonconvectivetwo-temperaturePCRamplification3PCRPCR。PCR751744a。4b。4Fig4Relationbetweendatacollectionroundandsimilarityoftopologyinferencein180nodenetwork4。。1DuffieldN．NetworktomographyofbinarynetworkperformancecharacteristicsJ．IEEETransactionsonInformationTheory200652125373－5388．2TianH．ShenH．Hammingdistanceandhopcountbasedclassifi-cationformulticastnetworktopologyinferenceC∥19thInterna-tionalConferenceonAdvancedInformationNetworkingandAp-plicationsTaipeiTaiwan2005267－272．3．J．20081082－86．4LiYongjunCaiWandongTianGuangli．Losstomographyinwire-lesssensornetworkusingGibbssamplingC∥ProceedingsofEWSN2007DelftNetherlands2007150－162．5IntanagonwiwatCGovindanREstrinD．DirecteddiffusionforwirelesssensornetworkingJ．IEEETransonNetworking20031112－16．6DownardI．SimulatingsensornetworksinNS—2R．TechnicalReportNRL/FR/5522—04—10073WashingtonDCUSANavalResearchLaboratory20041－9．1981－檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸檸。71。PCR23PCR、。、PCR。1．PCR/J．20052411－3．2．PCR/J．200423108－1013．3HashimotoMChenPCMitchellMWetal．RapidPCRinacontinuousflowdeviceJ．LabonaChip20044638－645．4KrishnanMUgazVMBurnsMA．PCRinaRayleigh-BénardconvectioncellJ．Science2002298793．5KrishnanMAgrawalNBurnsMAetal．ReactionsandfluidicsinminiaturizednaturalconvectionsystemsJ．AnalyticalChe-mistry2004766254－6265．6ChenZQianSAbramsWRetal．Thermosiphon-basedPCRreactorExperimentandmodelingJ．AnalyticalChemistry2004763707－3715．7WheelerEKBenettWStrattonPetal．Convectivelydrivenpo-lymerasechainreactionthermalcyclerJ．AnalyticalChemistry2004764011－4016．8AgrawalNHassanYAUgazVM．Apocket-sizedconvectivePCRthermocyclerJ．AngewandteChemieInternationalEdi-tion2007464316－4319．9ChungKHParkSHChoiYH．ApalmtopPCRsystemwithadisposablepolymerchipoperatedbythethermosiphoneffectJ．LabonaChip201010202－210．10LiuJinfengXingDaShenXingyanetal．DetectionofgeneticallymodifiedorganismsbyelectrochemiluminescencePCRmethodJ．BiosensorsandBioelectronics200420436－441．11ZhuDebinXingDaShenXingyanetal．HighsensitiveapproachforpointmutationdetectionbasedonelectrochemiluminescenceJ．BiosensorsandBioelectronics200420448－453．12ZhouXiaomingXingDaZhuDebinetal．MagneticbeadandnanoparticlebasedelectrochemiluminescenceamplificationassayfordirectandsensitivemeasuringoftelomeraseactivityJ．Ana-lyticalChemistry200981255－261．1974－MEMS。57