Abstract
The diethylaminoethyl (DEAE) dextran method for introducing cloned genes into mammalian cells is an alternative to the CaPO 4 technique for transformation. It has been observed that the DEAE dextran method is much better for studying the transient expression of transfected genes. In addition, it works better than CaPO 4 for some cell lines and is somewhat simpler to perform. Some cells, however, yield better results with CaPO 4 . The methods differ for nonadherent and adherent cells. The DEAE dextran method uses the cell-culture growth medium for cells to be transfected. The cell-growth medium consists of 250μg/ml DEAE dextran and 50 mM Tris, pH 7.4, with addition of 0.125 g DEAE dextran and 25 ml of 1 μ Tris, pH 7.4, to 500 ml of serum-free growth medium. The DEAE dextran method involves the harvesting of cells and analyzing them for a transient expression of the transfected gene. The cells are replated in a selective medium to obtain stable transfectants.
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