Production of purified recombinant virus samples, including cells and supernatants from plaque-purified recombinants are analyzed by Qbiogene or sent to the customer for analysis. Appropriate recombinants will be used for additional services. This service is performed using Sf9 cells grown in serum free medium. Time required: 5 weeks.
Infection of 4x10 6 Sf9 cells with three positive recombinant viruses
- Ethanol precipitation of transfer vector DNA for sterilization and quality control by agarose gel electrophoresis.
- Co-transfection of Sf9 insect cells with the recombinant transfer vector DNA and linear BacTenT baculovirus DNA.
- Isolation and plaque purification of 10 recombinant BacTenT viruses from the supernatant.
- Amplification by cell infection with each recombinant and preparation of viral DNA samples.
- Viral DNA analysis to ensure that the gene of interest is correctly introduced into the p10 locus of BacTenT.
50 µg of highly purified, verified transfer vector bearing the gene(s) to be expressed.
- 2 ml of each recombinant virus supernatant, as viral stocks.
- 1 ml of each supernatant to allow detection of secreted protein (if appropriate).
- 4x10 6 infected cells for the detection of the recombinant protein.
Next step : Plaque purification: Plaque purification of a recombinant Baculovirus prior to amplification