rt-pcr（rt - pcr）

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1、rt-pcr（rt-pcr）I.Experimentalapparatusandmaterials:1,transferguns:1ml,200L,20L,10L,2L2,suctionhead:1ml,200L,20L3,homogenatetube:5ml4,suctionheadtable:put1mlsuctionheadone,put20lsuctionheadone5,EPtubes:1.5ml,0.2ml,100L6,reagentbottle:260mlBrownreagentbo

2、ttle(widemouth,withcover)1125mlwhitereagentbottles(withabsoluteethanol)7,50ml,250ml,500ml:a8,capacitybottle:250ml,500ml,1000ml9,testtuberack:5ml,1.5ml,20L10,saltwaterbottles:250ml,500mleach2spare,onewithabsoluteethanol,anotherwithDEPCwater11,aluminuml

3、unchbox:412,plasticsmalllunchbox:113,largeporcelaincylinder:214,tinpaper:aroll15rollsofpaper:2rolls16,triangleflask:withacap,slightlylargerTwo,theprocessingandpreparationofexperimentalequipment1,plasticproducts:(includinggunhead,EPtube,homogenatetube,

4、etc.)TheDEPCofwaterfromtheflaskintotheceramiccylinder,theplasticproductsbysoakingthem,whichrequiresasmalltipStrawDEPCintothewater,andthendriedovernight,highpressure,spare,beforetheexperimentwillfirstputthegunsuctionhead,andahighpressure(EPtube)2,glass

5、products:acidsoakingovernight,washedclean,drysparefoilMongolia(DEPCblister)(washafterthefirstbubble1perthousandDEPCovernight,andthendried)3,homogenizer:(includingscissors,tweezers)firstwash,andthenhighpressure(donotneedtobubbleDEPC)Three.Reagentprepar

6、ation:1,DEPCwater:suck1ml,placedin1000mldoublesteamedwater,with1perthousandDEPCwater,placedinthe1000mlcapacitybottle,static4hoursstandby.2,75%ethanol:withanhydrousethanolDEPCwater,andthenput-20degreespreservation(whereDEPCwatershouldbehighpressure)3,i

7、sopropylalcohol:putbrownbottle4,chloroform:putinbrownbottle5agaroseFour.Preparationofseveralbuffers:1Electrophoresisbuffer:Tris54GBoricacid27.5g0.5M,EDTA,20ml?PH8.0Distilledwater1000ml5xTBE(ZhuCunye)The5xTBEisdiluted10timesto0.5TBE,whichcanbeusedinele

8、ctrophoresis(i.e.,theconcentrationoftheworkingfluid),suchas50mlstoragesolution,450mlwater,500mlworkingbuffer2,thesamplebuffer:0.25%bromophenolblue0.25%xylenegreenFF30%glycerin6*buffersolution,storedat4DEGCFive.Preparationofagarosegel:1and1%:1.