Sean Lauber:Adenovirus infection (in vitro)

This procedure is optimized for using 96-well plates (0.32 cm^2) with LLC cells. Find out how many cells are ideal for seeding for the time period you wish to study. In the case of LLC cells, 2000 cells/well is good for 48-72 hours.

1. Seed 2000 cells per well

2. Incubate overnight at 37°C/5% CO2.

3. Wash cells with 100 ul of PBS two times.

4. Add the appropriate pfu/cell (MOI) of the virus in 50 ul of PBS on top of the cells.

5. Incubate at 37°C for 30 minutes.

6. Add 50 ul of 2x media on top of the cells (don't remove the virus) - you want this media to contain 2x FBS.

7. Incubate for as long as you need at 37°C/5% CO2 (48-72 hours).

Possible to go from 20-50 pfu as well, although 10 seems to work just fine. Consider doing a dose response (MOI = 1, 10, 50).

PFU = MOI * #cells