High efficiency gene transfer by multiple transfection protocol.
A high efficiency transfection protocol employing a common polycationic lipid is described. Using LipofectAMINE, a widely used transfection reagent, we transfected 293T cells with a plasmid harboring the beta-galactosidase (beta-gal) gene. The transfection efficiency was determined by direct staining for X-gal. The conventional transfection protocol achieved an efficiency of <40% while our protocol, which employs the repetition of transfection a few times, achieved an efficiency of approximately 80%. Thus, a dramatic increase in transfection efficiency can be obtained by simply repeating transfection with the use of a common polycationic lipid. This method will be useful in many molecular biological experiments.
Yamamoto
M
M
Cardiovascular & Pulmonary Research Institute, Allegheny University of the Health Sciences, Pittsburgh, PA 15212, USA.
Okumura
S
S
Schwencke
C
C
Sadoshima
J
J
Ishikawa
Y
Y
D000818:Animals; D019556:COS Cells; D002411:Cation Exchange Resins; D002478:Cells, Cultured; D004337:Drug Carriers; D018014:Gene Transfer Techniques; D008055:Lipids; D014162:Transfection / Q000187:drug effects / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics*
D002411:Cation Exchange Resins; D004337:Drug Carriers; D008055:Lipids; C086724:Lipofectamine; D001616:beta-Galactosidase