adeasy 使用说明

AdEasy™AdenoviralVectorSystemINSTRUCTIONMANUALCatalog#240009(AdEasy™AdenoviralVectorSystem)#240005(pAdEasy-1vector)#240006(pShuttlevector)#240007(pShuttle-CMVvector)#240008(pShuttle-CMV-lacZ)#240081(pShuttle-IRES-hrGFP-1vector)#240082(pShuttle-IRES-hrGFP-2vector)RevisionA.01ForInVitroUseOnly240009-12LIMITEDPRODUCTWARRANTYThiswarrantylimitsourliabilitytoreplacementofthisproduct.Nootherwarrantiesofanykind,expressorimplied,includingwithoutlimitation,impliedwarrantiesofmerchantabilityorfitnessforaparticularpurpose,areprovidedbyAgilent.Agilentshallhavenoliabilityforanydirect,indirect,consequential,orincidentaldamagesarisingoutoftheuse,theresultsofuse,ortheinabilitytousethisproduct.ORDERINGINFORMATIONANDTECHNICALSERVICESUnitedStatesandCanadaAgilentTechnologiesStratageneProductsDivision11011NorthTorreyPinesRoadLaJolla,CA92037Telephone[phone]OrderTollFree(800)424-5444TechnicalServices(800)[e-mail](0)20312570008000230448008007000700000800700170010080074007400Switzerland080056308008005630820800563081008007000700000800700170010080074007400UnitedKingdom080091732820800917328308009173281AllOtherCountriesPleasecontactyourlocaldistributor.Acompletelistofdistributorsisavailableat™AdenoviralVectorSystemCONTENTSMaterialsProvided..............................................................................................................................1StorageConditions..............................................................................................................................1AdditionalMaterialsRequired..........................................................................................................2NoticestoPurchaser...........................................................................................................................3Introduction.........................................................................................................................................7OverviewoftheAdEasy™System.......................................................................................8AdvantagesofRecombinantAdenovirusforGeneExpression............................................8VectorFeatures....................................................................................................................10pAdEasy-1VectorMap.......................................................................................................11pShuttleVectorMap............................................................................................................12pShuttle-CMVVectorMap.................................................................................................13pShuttle-IRES-hrGFP-1VectorMap..................................................................................14pShuttle-IRES-hrGFP-2VectorMap..................................................................................15pShuttle-CMV-lacZVectorMap.........................................................................................16RecommendedPrimerSequences.......................................................................................17FeaturesofBacterialStrains............................................................................................................17GeneratingAdEasy™Recombinants..............................................................................................18CloningConsiderations.......................................................................................................18CloningtheGeneofInterest................................................................................................20TransformationGuidelinesforBJ5183Cells......................................................................20CotransformingtheBJ5183CellstoProduceRecombinantAdPlasmid...........................21TestingColoniesforRecombinantAdPlasmid..................................................................22InterpretationofResults......................................................................................................23AmplifyingRecombinantAdPlasmid.............................................................................................24TransformationGuidelinesforXL10-GoldCells...............................................................24XL10-GoldTransformationProtocol..................................................................................25PreparationofPrimaryAdenovirusStockwithRecombinantAdPlasmid................................27SafetyConsiderations..........................................