Transfection by Electroporation of RAW 264.7 Macrophages
Adapted from Transcriptional Regulation in Eukaryotes: Concepts, Strategies, and Techniques, 2nd edition, by Michael F. Carey, Craig L. Peterson, and Stephen T. Smale. CSHL Press, Cold Spring Harbor, NY, USA, 2008.INTRODUCTION
In electroporation-based transfection methods, the DNA and cells are mixed in an empirically defined buffer, which establishes the resistance during the electric shock. The solution is placed in a cuvette that is compatible with the electroporation device being used. The mixture is then subjected to a brief high-voltage shock, which induces or stabilizes pores in the plasma membrane through which the DNA enters the cell. The cells are allowed to recover by a brief incubation at room temperature and then transferred to a tissue culture plate or T-flask along with growth medium and serum. Electroporation is an attractive method for transfection because it is more rapid and involves fewer manipulations than other methods and can be used for many types of adherent and nonadherent cells. This protocol describes transfection by electroporation of RAW 264.7 macrophages.
