Lipofectamine plasmid transfection

For one reaction, 2 µg of DNA plasmid was mixed with 5 µl of Lipofectamine 2000 (#11668019; Thermo Fisher Scientific) in serum-free medium to a total volume of 200 µl and incubated at room temperature for 5 min before adding to 3 × 10⁵ cells in a six-well plate. Cells were incubated at 37°C, 5% CO₂, and the transfection efficiency was assessed after at least 18 h.