Summary of Optimal Transfection Conditions: Confluence on the day of transfection Cell culture conditions GenJet™ (µl) : DNA (µg) Ratio Diluent for DNA and Transfection Reagent Incubation Time to Form GenJet™/DNA Complex Presence of Serum/Antibiotics during Transfection Change Medium 5 Hours After Transfection Maximal Efficiency Transfection Results: Reporter Gene Plasmid Efficiency (GFP %) | ~80% DMEM with 4.5 g/L glucose, 10% FBS 3:1 Serum-free DMEM with 4.5 g/L glucose 15 minutes at RT Yes Yes 48 hours EGFP pEGFP-N3 (CMV promoter) 88% |