磷酸钙转染方法的优化

The Optimization of Calcium Phosphate Transfection Method

通过比较293T细胞的接种密度、质粒DNA的量以及CO2浓度对磷酸钙转染方法的影响,确定最佳的转染条件.以各单因素试验的最适条件为依据,设计三因素三水平的正交试验.结果表明,磷酸钙介导质粒DNA转染,当293T细胞的接种密度为5×105/mL、质粒DNA的量为30μg、细胞培养环境CO2浓度为3%时,转染效果最佳.

In this study,the effects of seeding density of 293T cells,as well as the amount of plasmid DNA and the concentration of CO2 in culture environment,on the efficiency of calcium phosphate transfec—tion were compared,optimum transfection condition was confirmed.On the basis of optimum condition obtained from single factor experiments,orthogonal tests in three levels of three factors were designed and performed.The results showed that the highest transfection efficiency was obtained when the seeding den—sity of 293T cells was 5×10^5/mL,the amount of plasmid DNA was 30μg and the concentration of CO2 in culture environment was 3%.