Transduction and titration of lentiviral supernatant

Materials
	DMEM + 10% CS
	Puromycin (stock solution 5 mg/ml)

Procedure

1. 1x10⁴ 293T cells are seeded in a 24-well plate ~12 h before transduction (5 wells)
2. 1µl, 2µl or 5µl, respectively, of the virus-concentrate are added on top of the cells, 2 wells remain untreated
3. incubation over night at 37°C and 5% CO₂
4. 24 h after transduction the medium is replaced by 1 ml fresh medium containing 10% CS and 0,4 µg/ml puromycin (one of the wells harbouring untransduced cells will not be treated with puromycin)
5. after 4-5 days the cell number is defined and the viral titer (viral particles / µl) is calculated by counting still viable cells