Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Temperature: 37°C
Recommendations for Infection: Plate cells 16-24 hours prior to infection and infect when cultures are 80-90% confluent. Remove medium and inoculate with a small volume of virus (e.g. 1 mL per 25 cm2) diluted to provide an optimal MOI (e.g. 0.01). Adsorb 1-2 hours at 37°C in a humidified x% CO2 atmosphere. End adsorption by adding virus growth medium. Replace virus growth medium at one day post-inoculation to remove DMSO from the culture.
Harvest by removing culture medium and adding cell dissociation buffer. Dilute in freeze medium containing a final concentration of 7% DMSO and preserve with a controlled-rate freeze (1°C/minute).
Incubation: 3-4 days at 37°C in a humidified 5% CO2 atmosphere, until CPE is progressed through 60% of the monolayer.
Takahashi M, et al. Development of a live attenuated varicella vaccine. Biken J. 18: 25-33, 1975. PubMed: 167707
