Co-transfection → recombinant virus
Construction of the recombinant baculovirus via homologous recombination relies on the success of the co-transfection. OET offers a cost-effective approach to customers who wish to ensure success with their baculovirus co-transfection(s). For the co-transfection reaction, a transfer plasmid containing the gene of interest needs to be either provided by the customer or produced by OET. This transfer plasmid is then used with the flashBAC ULTRA baculoviral DNA to co-transfect insect cells (Sf9). A control co-transfection is also performed in which a transfer plasmid containing the lacZ gene is used. Following incubation of cells for five days at 28°C, the cell culture medium containing the recombinant virus is harvested. This virus stock (P0) is then sent to the customer.
- Gene synthesis and gene cloning
- Virus stock amplification
- Titration of virus by plaque assay
- Titration of virus using qPCR (baculoQUANT)