Cell-free BoDV preparation.

Thank you for your interest in our research. The following are the details of the protocol you asked for.
 

1. Prepare BoDV-infected cells (OL or Vero) in 15-cm dish
2. Wash the cells with 10 mL of 20 mM HEPES buffer (pH 7.4) twice
3. Incubate the cells with 5 mL of 20 mM HEPES buffer (pH 7.4) containing 250 mM MgCl2 and 1% FCS for 90 min at 37°C
4. Collect the culture supernatant
5. Centrifuge the culture supernatant at 3000 rpm for 5min at 4°C
6. Filter the supernatant through 0.22 µm membrane filters
7. Filtered solution should be stored at -80°C