A low-titer solution from a single recombinant is used to generate a titrated high-titer stock. This service is performed using Sf9 cells grown in serum free medium. Time required: 3 to 4 weeks.
- Infection of 1.2x107 Sf9 cells in a 75 cm2 flask and harvesting of supernatant at 72 hours post-infection (p.i.).
- Second round of infection in flasks and harvesting of supernatant at 72 hours p.i.
- Third round of infection or in spinners and harvesting of supernatant at 72 hours p.i.
- Viral titer determination using plaque assay.
2 ml of a low-titer stock, preferably with a known viral titer.
100 ml, 500 ml or 1 liter of recombinant virus stock at approximately 108 pfu/ml.
Next step: Purification: Sucrose gradient purification of a recombinant baculovirus
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