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: For best results, infection should be performed on a 90-100% confluent, 18-48 hour old cellular monolayer. Activate virus by incubation with 10 µg/mL trypsin type IX for 60 minutes at 37°C. Prepare dilution of virus to no greater than 2 µg/mL trypsin in minimum amount of volume (e.g. 1 mL per 25 cm2). Wash monolayer three times with PBS or serum free medium prior to inoculation. Adsorb virus dilution for 1 hour at 37°C in a humidified 5% CO2 atmosphere, rocking every 20-30 minutes to redistribute inoculum. End adsorption by removing the inoculum, washing the monolayer once with PBS or serum free media, and then adding virus growth medium.
Incubation: 2-3 days at 37°C in a humidified 5% CO2 atmosphere, until CPE is progressed through 80% of the monolayer.
Arnold M, Patton JT, McDonald SM. Culturing, storage, and quantification of rotaviruses. Curr. Protoc. Microbiol. Chapter 15: Unit 15C.3, 2009. PubMed: 19885940
Offit PA, Clark HF, and Plotkin SA. Response of mice to rotaviruses of bovine or primate origin assessed by radioimmunoassay, radioimmunoprecipitation, and plaque reduction neutralization. Infect. Immun.42(1): 293-300, 1983. PubMed: 6311747
