Recommendations for Infection: Plate cells 24-48 hours prior to infection and infect when cultures are 80-90% confluent. Remove medium and then wash monolayer with PBS or serum free medium prior to inoculation. Inoculate with a small volume of inoculum (e.g. 1 mL per 25 cm2) diluted to provide an optimal MOI (e.g. 0.01). Adsorb for 1 hour at 33°C in a humidified 5% CO2 atmosphere. End adsorption by adding virus growth medium.
Incubation: 3 days at 33°C in a humidified 5% CO2 atmosphere while rocking, until CPE is progressed throughout