Investigation on optimal parameters of electroporation-mediated transfection in 32DP210

Objective:To find the best parameters to transfect plasmid pG into 32DP210 cells.Methods:LipofectamineTM2000,transfast,liposome DMRIC-E transfection and electroporation were compared for transfecting plasmid pG,a plasmid that expressed G418 resistance protein and green fluorescent protein(GFP),into 32DP210 cells.The expression of GFP was observed and the transient transfection efficiency was measured after the transfection by fluorescence microscope.The stable transfected cells were screened by G418.Results:The optimum conditions for laboratory electroporation-mediated 32DP210 cell transfection were:power transfer voltage 270 V,capacitance of 950 μF,DNA dose of 14 μg.Transfection efficiency of electroporation after 48 h was 50.81%,while the other three methods were less than 3%.Conclusion:Electroporation is the best way to stably transfect plasmid pG into 32DP210 cells.