Transfection of Primary Rat Cortical Cultures with 
Tfx^™-50 Reagent: Optimization of Gene Expression

by Faye A. Boeckman, Rachel Margaret Whelan and Karen Hartnett 
Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261 USA

Traditional methods of transfecting primary cell cultures, such as calcium phosphate precipitation and electroporation, typically result in few transformants. Lipid-mediated gene transfer, however, is successful for many different cell types including primary cultures. In this report, we describe the optimization of transfection using Promega’s Tfx^™-50 Reagent (Cat.# E1811), and compare the transfection efficiency of this reagent to that of the polycationic lipid 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-
N,N-dimethyl-1- propanaminium trifluoroacetate (DOSPA). Our neuronal cell culture model consists of primary embryonic rat cortical cells. In addition, we determine the duration of expression with the reporter genes luciferase and beta-galactosidase as transfection markers.

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