Recommendations for Infection: Plate cells 24-48 hours prior to infection and infect when cultures are 90% confluent. Remove CGM and then wash monolayer once with 1X DPBS (ATCC® 30-2200™). Inoculate with a small volume of virus (e.g. 1 mL per 25 cm2) diluted in plain DMEM to provide an optimal MOI (e.g. 0.001). Adsorb 2-2.5 hours at 32°C in a humidified 5% CO2 atmosphere, rotating on a roller apparatus at 0.75 RPM. End adsorption by adding virus growth medium.
Incubation: Incubate infected culture for 21 days at 32°C in a humidified 5% CO2 atmosphere on a roller apparatus at 0.75 RPM.