Baculovirus --- infection

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Adherent culture

1. Seed cells in tissue culture dishes or flasks (e.g., 106 cells in 35 mm dish).

2. Let cells attach for 1 hour or overnight.

3. Aspirate media and replace with 0.2 ml (35 mm dish) fresh medium containing viruses (1-5 MOI).

4. Incubate at room temperature for 1 hour.

5. Remove medium and replace with fresh medium.

6. Incubate at room temperature or 27 °C.

7. Harvest for protein 36-72 hours after infection. Harvest for viruses 2-6 days after infection.

Suspension culture

1. Dilute cells to ~0.5x106 cells/ml in fresh medium.

2. Infect with virus (0.1-5 MOI).

3. Incubate at room temperature or 27 °C on a roller or with shaking.

4. Harvest for protein 2-3 days after infection. Harvest for viruses 2-6 days after infection.