关键词: Lipofectamine 3000, FuGENE HD, DNA-In CRISPR, pAdeasy-1-EGFP, SKOV3细胞, 大片段质粒

Abstract: The plasmid DNA pAdeasy-1 EGFP, coding for green fluorescent protein, was transfected into SKOV3 cells via 3 different commercial agents (Lipofectamine 3000, DNA-In CRISPR and FuGENE HD, respectively). We examined the transfection efficiency of reagents to choose the appropriate transfection reagent for solving the problems of large fragment plasmid with lower transfection efficiency. Transfection efficiency was determined by green fluorescent protein (GFP) fluorescence using fluorescence microscope, and cell activity was assessed by cell counting kit-8 (CCK-8). The results showed that the transfection efficiency of Lipofectamine 3000 was more than 50%, the efficiency of FuGENE HD was about 20%, and that of the DNA-In CRISPR was less than 5%. After 24 h transfection, the cell activities with Lipofectamine 3000, FuGENE and DNA-In CRISPR were （97.5±2.52）%,（89.5±2.43）% and (95.5±3.48）%, respectively. Comparative analysis showed Lipofectamine 3000 had relatively high transfection efficiency with minimal cytotoxicity, indicating that Lipofectamine 3000 could be a proper choice for transporting massive plasmid into SKOV3 cells.

Key words: Lipofectamine 3000, FuGENE HD, DNA-In CRISPR, pAdeasy-1-EGFP, SKOV3 cells, large fragment plasmid