High Efficiency Gene Transfer by Multiple Transfection Protocol
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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 β-galactosidase (β-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.
KeywordsLipid Gene Transfer 293T Cell Transfection Efficiency Transfection Reagent
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- Cannon SC, Strittmatter SM (1993) Functional expression of sodium channel mutations identified in families with periodic paralysis. Neuron 10: 317-326.Google Scholar
- Edlund T, Walker MD, Barr PJ, Rutter WJ (1985) Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5′ flanking elements. Science 230: 912-916.Google Scholar
- Felgner PL (1993) LipofectAMINE™ reagent: a new, high efficiency polycationic liposome transfection reagent. Focus 15: 73-79.Google Scholar
- Hawley-Nelson P, Ciccarone V, Gebeyehu G, Jessee J, Lojda Z (1970) Indigogenic methods for glycosidases. Histochemie 23: 266-288.Google Scholar
- Maniatis T, Fritsch EF, Sambrook J (1989) In: Ford N, Nolan C, Ferguson M, Ockler M, eds. Molecular Cloning: A Laboratory Manual. 16.45. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
- Russell DW, Miller AD, Alexander IF (1994) Adeno-associated virus vectors preferentially transduce cells in S phase. Proc Natl Acad Sci USA 91: 8915-8919.Google Scholar