User:Karmella Haynes/Notebook/miR Trigger/2009/12/04

Project name

Main project page Previous entry      Next entry

12/04/09 Transfection: KAH93, 94, 95/MV2 (Plasmids not yet confirmed, postpone) Transfections > U2OS cells + Gal4DB-PEST-RFP constructs > Use Lipofectamine, 6-well format Wells Plasmid DNA Volume Fugene Opti-MEM (total) 1 KAH93/MV2 2 μg ## μL 4 μL 500 μL 2 KAH94/MV2 " ## μL " " 3 KAH95/MV2 " ## μL " " 4 KAH36(control) " ## μL " " 5 mock --- --- " " 6 untreated --- --- --- " > Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min. > Add DNA to 250 μL Opti-MEM > Add DNA mix to Lipo mix --> R.T./ 20 min. > Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37°C > Assay RFP after 18-24 hours