Recommendations for Infection: For best results, infection should be performed on an 80-90% confluent, 18-48 hour old cellular monolayer. Prepare dilution of virus in minimum amount of volume (e.g. 1 mL per 25 cm2). Wash monolayer with PBS or serum free medium prior to inoculation. Adsorb virus dilution for 1-2 hours at 37°C in a humidified 5% CO2 atmosphere, rocking every 20-30 minutes to redistribute inoculum. End adsorption by adding virus growth medium.
Incubation: 3-7 days at 37°C in a humidified 5% CO2 atmosphere, until CPE is progressed through 80% of the monolayer.