Recommendations for Infection: Plant cells 24 hours in advance and infect when cultures are 90-100% confluent. Remove medium and inoculate with an appropriate volume of Rickettsiae (e.g. 1 mL per 25 cm2) diluted in agent growth medium to provide an appropriate MOI (e.g. MOI of 50 for 50 Rickettsiae/ cell). Adsorb 1 hour at 800 x g at 20°C in a low speed centrifuge in closed buckets. End adsorption by adding agent growth medium to culture(s).
Incubation: Incubate infected culture(s) for 7-14 days at 35°C in a humidified 5% CO2 atmosphere.