Optimization of Cell Transfection Conditions
LIU Yun-hai,ZHONG Ying-li,REN Kai-qun,ZHOU Jian-lin,ZHANG Jian,Han Mei(Department of Biochemistry and Molecular Biology,Life Science College of Hunan Normal University,Changsha 410081,China)
Perfect cell transfection efficiency was acquired with the comparisons of three cell transfection assays(Electroporation, calcium phosphate method and lipofectin reagent) to carry out further studies on gene expression regulation and mouse ES cell targeting. The transfection efficiency of electroporation was linear with the voltage which was set from 350 to 450 V; the transfection efficiency of calcium phosphate was greatly improved by using HEPES buffer; poor quality of DNA and longer incubating time were associated with decreased transfection efficiency of lipofectin. According to the characteristics of the three transfection protocols, transfection protocol can be optimized to obtain higher efficiency.