JetPEI transfection

JetPEI (Polyplus Transfection cat#PP10140N)

- Set up two separate mixes: for 10cm (for 6w)

Mix A) 150 mM NaCl (sterile) 250 µL 42 µL

DNA 6 µg (*) 1 µg

Mix B) 150 mM NaCl (sterile) 250 µL 42 µL

JetPEI 12 µL (*) 2 µL (JetPEI must be double the amount of DNA)

- IMPORTANT: add Mix B to Mix A (not the reverse!!) = 500 µL (84 µL)

- Leave at RT for 20 minutes

- In the meantime, wash cells with PBS (to get rid of dead cells)

- Add 6 ml (1 mL) of complete medium to the cells (it doesn't matter if it contains FCS)

- Add Mix A+B directly to the plate, drop-wise, while swirling the plate

- Change the medium after 6 hrs or the day after

(*) (**) It is possible to use twice as much DNA (12 and 2 µg), but then you should also double the amount of JetPEI (24 and 4 µL, respectively)