Effect of some factors on transformation of mammalian cells with exogenous DNA

Objective:To set up a standard method of gene transfection in our laboratory.Methods:Using the calcium phosphate-DNA coprecipitation technique,mammalian cell line (HeLa)were transformed with several kinds of the recombinant plasmid DNA.The effect of some factors on transformation efficiency was further observed.Results:The host cells for transfection should be of growth at the log phase;The concentration of the recombinant plasmid DNA is about 8 to 10 μg per flask;The incubation time between DNA-phosphate coprecipitation and HeLa cell is 10 hours;Antibiotic G418 was treated as a concentration of 200 μg/mL for selective culture,and added 24 hours after transformation.Conclusion:These findings will provide the favorable conditions for the high level expression of exogenous gene in vitro.