Low transfection rate of electroporation

[chenln]

Hi, there,

I am trying electroporate cos-7 using eppendorf's multiporator but get poor transfection rate. The survival rate is 40% and transfection rate(based on the survived cell number) is 40%. So I get only 16% cell transfected based on initial cell number. My transfection varibles is 400V, 100us, 200ul, 0.2 cm gap cuvette, hyposmotic solution, room temperature. But eppendorf claims that 30-40% cell transfected based on initial cell number. The recommended conditions are 600V, 40us, 400ul, 0.2 cm gap cuvette, hyposmotic solution, on ice. But by following this, I got very poor survival rate(almost 1%). I found at room temperature, the survival rate is better keeping other conditions same, but transfection rate based on survived cells is only 20%. 

Is there any one can give me some suggestions?


Thanks,

Dr. Chen linan

Department of Anatomy
Beijing Medical university


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