One hour prior to transfection, the culture medium was replaced with serum-free RPMI medium. 0.4 μg of DNA (pNN9 or DNA ministring), diluted in 50 μl of serum-free OptiMEM culture medium, was mixed with different aliquots of 1.5 mM 16-3-16 gemini surfactant solution to yield N + /P − charge ratios of 3:1 and 5:1.
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