Forresearchuseonly.Notforhumanoranimaltherapeuticordiagnosticuse.Lipofectamine®RNAiMAXReagentCat.nos.13778-100Size:0.1mLStoreat4°C(donotfreeze)13778-0300.3mL13778-0750.75mL13778-1501.5mL13778-50015mLPub.PartNo.13778.ppsPub.No.MAN0001182Rev.Date20July2012Description•Lipofectamine®RNAiMAXTransfectionReagentisaproprietaryformulationfortransfectingsmallRNAs(e.g.,siRNA,Silencer®SelectsiRNA,Stealth®RNAi,mirVana™miRNAmimicsandinhibitors)intoawiderangeofeukaryoticcells.•Nucleicacid-Lipofectamine®RNAiMAXcomplexescanbeaddeddirectlytocellsinculturemedium,inthepresenceorabsenceofserum.•Itisnotnecessarytoremovecomplexesorchange/addmediumaftertransfection.ImportantGuidelinesforTransfection•UseOpti-MEM®IReducedSerumMedium(Cat.no.31985-062)todiluteLipofectamine®RNAiMAXTransfectionReagentandRNA.•ResuspendsiRNAwithnuclease-freewateratarecommendedstockconcentrationof100μM.Storealiquotsat–20°C.•DilutestocksiRNAtoaworkingconcentrationof10μMfortrans-fection.A10μMstockofsiRNAduplexisequivalentto10pmol/μL.•Use30nMRNAiduplexasastartingpoint.BLOCK-iT™AlexaFluor®RedFluorescentOligo(Cat.no.14750100)canbeusedtodeterminetransfectionefficiency.•Foradditionalinformationandprotocols,refertothemanualatwww.lifetechnologies.com/transfection.CellsLipofectamine®RNAiMAXReagentsiRNAAdddilutedsiRNAtodilutedLipofectamine®RNAiMaxReagent(1:1ratio).DiluteLipofectamine®RNAiMAXReagentinOpti-MEM®Medium.DilutesiRNAinOpti-MEM®Medium.Incubate.Seedcellssotheywillbe60–80%confluentatthetimeoftransfection.Visualize/analyzetransfectedcells.AddsiRNA-reagentcomplextocells.Day0Day1Day2–45RNAiTransfectionTransfectcellsaccordingtothefollowingtable.Thetransfectionisdesignedfor1RNAamountcombinedwith1amountofLipofectamine®RNAiMAX.Thepreparedmixisenoughtohavetriplicates(96-well),duplicates(24-well),andsinglewell(6-well)transfections,andaccountforpipettingvariations.Component96-well24-well6-wellAdherentcells1–4x1040.5–2x1050.25–1x106Opti-MEM®Medium25µL50µL150µLLipofectamine®RNAiMAXReagen...
