Baculovirus stocks

Baculovirus stocks are typically produced by the aforementioned methods, followed by optional amplification rounds and titration. The traditional titration technology is based on plaque assays where viral stock dilutions are applied on confluent insect cell cultures and clonal plaques scored by visual inspection after several days. More rapid technologies include 1) immunodetection of viral proteins ([9], BacPAK Rapid Titer Kit, BD Biosciences) 2) fluorescent... 

Protocol 1.9 Amplification of the recombinant Baculovirus virus stock... 

Seed Sfi cells at a density of approximately 60% (3.6 x 10 cells) and allow attaching to the cell culture flask (150 cm ) for 30 min. Aspirate the medium and add 5 mL of the virus-containing supernatant (step 9) together with 3 mL of Insect-Xpress to the freshly seeded Sfi cells. Incubate for 1 h at 27 °C. Then add 22 mL of Insect-Xpress supplemented with 10% PCS and 50pg/mL gentamicin and continue the incubation for 5—7 days (second round of amplification). High-titer stocks of the baculoviruses are obtained by at least three cycles of amplification in Sfi cells. 

The supernatant containing recombinant baculovirus was transferred to a fresh tube and stored at 4°C this is the stock. 

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