STEWART LABORATORY

In Vitro AdCre INFECTION

Plate the cells at a relatively low density, and the infection should be performed in a minimal amount of media. This protocol is for a 6 well plate,
but you can scale up or down as needed

1. Trypsanize cells; quench

2. Spin down the cells and re-suspend in a very small amount of infection media (you can dilute later if necessary); count the cells

3. Dilute with infection media so that the concentration is about 2 x 10⁵ cells/mL

4. For each well, use 500 uL of cells. Add the appropriate amount of virus and mix well (Probably around 1-5 µL of high-titer stock – you want to get an MOI of around 10)

5. Plate cells and incubate for ~ 2 hrs at 37°C

6. Add 2 mL full media and let cells grow for ~ 48 hrs

Infection media: DME + Pen/Strep + 2% IFS + Glutamine

Full media: DME + Pen/Strep + 10% IFS + Glutamine