Lipofectamine2000转染说明书

Tip:1.Tosearchthenote,pressCTRL-Fkeysincombination2.Tonavigatethenote,usethelinks.Lipofectamine™2000TransfectionReagentWewanttohearfromyou!ClickhereTABLEOFCONTENTSPRODUCTDESCRIPTIONSHIPPINGCONDITIONSSTORAGECONDITIONSSTABILITYQCSPECIFICATIONSPROTOCOL&APPLICATIONNOTESRelativeSurfaceAreasOfTissueCultureVesselsSurfaceAreasOfTissueCultureVesselsTubesRecommendedForUseWithLipofectamine™2000OptimizingPlasmidDNATransfectionsWithLipofectamine™2000TransfectionProtocolsCo-TransfectionOfSirnaAndPlasmidDNATransfectionOfFluorescentlyLabeledOligosWithLipofectamine™2000TransfectionOfCellsIn96-WellPlatesTransientTransfectionOfSuspensionCellsALTERNATEPRODUCTS&COMPATIBILITYPRODUCTDOCUMENTATIONREFERENCESPRODUCTNAME&CATALOGNUMBERSASSOCIATEDPRODUCTSRELATEDTECHNICALSUPPORTNOTES1PRODUCTDESCRIPTION(backtoTableofContent)Lipofectamine™2000TransfectionReagentisaproprietaryformulationforthetransfectionofnucleicacids(DNAandRNA)intoeukaryoticcellsandprovidesthefollowingadvantages:Highesttransfectionefficiencyinmanycelltypesandformats(e.g.96-well).RefertotheCellLinesDatabaseforalistofcelltypessuccessfullytransfected.Detailedin-housetransfectionprotocolsarealsoavailableatthissitewhereavailable.DNA-Lipofectamine™2000complexescanbeaddeddirectlytocellsinculturemedium,inthepresenceorabsenceofserum.Lipofectamine™2000maybeusedinthefollowingapplications:TransientandstabletransfectionofadherentandsuspensioncellsHighthroughputtransfectionsDeliveryofStealthRNAiandsiRNAintocellsForinformationontransfectingmammaliancellswithshortinterferingRNAs(siRNA)foruseinRNAinterference(RNAi)studies,visitourRNAiCentralwebpageat™2000givessuperiortransfectionefficiencyforthefollowingcelllines:293F293HBE(2)C(w/oserum)CHO-K1CHO-S(adherent)COS-1(w/oserum)COS7-LPrimaryHumanFibroblasts(w/oserum)HT-29(w/oserum)HT-1080MDCKMRC-5(w/oserum)PC12SK-BR3VeroHepG2NIH3T3Lipofectamine™2000CDisa100%syntheticversionofLipofectamine™2000.UseitinthesamewayasLipofectamine™2000,butbesuretouseanimalorigin-freereagents.SHIPPINGCONDITIONS(backtoTableofContent)Thiskitisshippedonwetice.STORAGECONDITIONS(backtoTableofContent)Lipofectamine™2000TransfectionReagentshouldbestoredat40C.STABILITY(backtoTableofContent)ThestabilityofLipofectamine™2000TransfectionReagentisguaranteedfor6monthswhenithasbeenstoredasrecommended.Lipofectamine™2000Reagentshouldnotbefrozen.QCSPECIFICATIONS(backtoTableofContent)Lipofectamine™2000istestedfortheabsenceofmicrobialcontaminationusingbloodagarplates,Sabarauddextroseagarplates,andfluidthioglycolatemedium,andfunctionallybytransfectionofCHO-K1cellswithareporterplasmid.PROTOCOLANDAPPLICATIONNOTES(backtoTableofContent)GeneralprotocolnotesSurfaceAreasOfTissueCultureVesselsTubesRecommendedForUseWithLipofectamine™20002OptimizingPlasmidDNATransfectionsWithLipofectamine™2000TransfectionProtocolsCo-TransfectionOfSirnaAndPlasmidDNATransfectionOfFluorescentlyLabeledOligosWithLipofectamine™2000TransfectionOfCellsIn96-WellPlatesTransientTransfectionOfSuspensionCellsGeneralprotocolnotes(backtoTableofContent)(backtoProtocolandApplicationNotes)Itisnotnecessarytoremovecomplexesorchange/addmediumaftertransfection,butcomplexesmayberemovedafter4-6hours.DMEMorRPMI1940canbeusedinsteadofOpti-MEMwhenmakingLipofectamine™2000–DNAcomplexes.However,theefficiencyofcomplexformationmaynotbeashighaswithOpti-MEM.CellsinPBScanbetransfectedusingLipofectamine™2000.AslongasthecellsarehealthyinthePBS,thetransfectionislikelytowork.ForageneralplasmidDNAtransfectionprotocol,pleaserefertotheproductinsert:™2000_man.pdfAgeneralprotocolfortransfectingStealthRNAiorsiRNAintomammaliancellscanbefoundatthefollowingsite:(backtoTableofContent)(backtoProtocolandApplicationNotes)CultureVessel96-well48-well24-well12-well6-well35-mm60-mm100-mm150-mmT25T75SurfaceArea(cm2)0.30.724101020601402575Ratioto24-wellplate0.20.4125510307012.537.5TubesrecommendedforusewithLipofectamine™2000(backtoTableofContent)(backtoProtocolandApplicationNotes)Itisbesttousepolypropylenetubeswhenpre-mixingLipofectamine2000™andDNA.Polystyrenemaynotworkaswell.OptimizingplasmidDNAtransfectionswithLipofectamine™2000(backtoTableofContent)(backtoProtocolandApplicationNotes)TheconditionsthatcouldbeoptimizedincludeLipofectamine™2000amount,DNAconcentration,andcellnumber.Keepingtwovariablesconstant,varythethird.Forexample:tooptimizetheamountofLipofectamine™2000fortransfectionina24-wellplate,startwithcellsat90%confluencyanduseafixedamountofDNA(0.8-1.2µg).WithcellnumberandDNAconcentrationheldconstant,varytheamountofLipofectamine™2000todeterminetheoptimalconcentration(usually1.5-3µl).Inthesameway,thecellnumberandamountofDNAcanalsobeoptimized.Itisrecommendedtousearangeof0.5to5µlofLipofectamine™2000perµgofDNA.Itispossibletominimizetheeffectoftransfectiononcellgrowthandviabilitybyincreasingthenumberofcellspl