预优化DNA转染试剂CV1相关交流-蚂蚁淘在线

预优化DNA转染试剂CV1

mayitao /发布时间:1548570213 /浏览量:69

Description

GenJet™DNAInVitroTranfectionReagentforCV-1ispre-optimizedfortransfectingCV-1cells. TheCV-1celllinewasinitiatedinMarchof1964withatissuesectionexcisedfromthekidneyofanadultmaleAfricangreenmonkey(Cercopithecusaethiops).Thefibroblastlinewasoriginallyutilizedinresearchfocusingonthetransformationofthecancer-causingRoussarcomavirus(RSV),butnowispopularasahostforacquiredimmunodeficiencydisease(AIDS)research,aswellastransfectionexperimentswithsimianvirus40(SV40)andrecombinantplasmidvectors.CV-1cellsexhibitfibroblastmorphology,growadherentlytoglassorplasticsurfaces,andarenegativeforreversetranscriptase.Thecellsareknowntobesusceptibletoseveralviruses,includingpoliovirus1,herpessimplex,simianvirus40,Californiaencephalitis,andbothEasternandWesternequineencephalitis.Thecellsexhibitrapidgrowthandchromosomenumbershiftshavebeenreportedtooccurathighpassagelevels.RefertothefollowingoptimaltransfectionconditionsformaximaltransfectionefficiencyonCV-1cells.GenJet™reagent,1.0ml,issufficientfor300to600transfectionsin24wellplatesor150to100transfectionsin6wellplates.

SummaryofOptimalTransfectionConditions:ConfluenceonthedayoftransfectionCellcultureconditionsGenJet™(µl):DNA(µg)RatioDiluentforDNAandTransfectionReagentIncubationTimetoFormGenJet™/DNAComplexPresenceofSerum/AntibioticsduringTransfectionChangeMedium5HoursAfterTransfectionMaximalEfficiency TransfectionResults:ReporterGenePlasmidEfficiency(GFP%)

~80%DMEMwith4.5g/Lglucose,10%FBS3:1Serum-freeDMEMwith4.5g/Lglucose15minutesatRTYesNo36hoursEGFPpEGFP-N3(CMVpromoter) 75%

StorageConditionStoreat4°C.Ifstoredproperly,theproductisstablefor12monthsorlongerAPictureShowingTransfectionEfficiencyofGenJet™ReagentonCV-1Cells. CV-1cellsweregrownperATCCrecommendedculturemediumandtransfectedwithpEGFP-N3byGenJet™.Theefficiencywaschecked24hoursposttransfection

DataSheet.pdf

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