Viruses Purification of Perkinsus spp.
First, Fill centrifuge bottles with 450mL Perkinsus Culture in the logarithmic growth phase.
Centrifuge bottles at 1500 g; 15 min at room temperature.
Transfer supernatant to a clean tube, and store at 4°C for further analyses.
Resuspend pellet in 10% initial culture volume supernatant in a 50mL Falcon Tube.
Centrifuge tubes at 1500 g; 15 min at room temperature.
Transfer supernatant to a clean tube, and store at 4°C for further analyses.
Resuspend pellet in 4mL PBS/0.35 M NaCl) containing protease inhibitor cocktail.
Transfer the pellet resuspended in 2mL Eppendorf Tubes (1.5mL/tube).
Add 100mL Glass beads.
Disruptor Genie for 15sec.
Verify cell disruption : place a 20mL under light microscope.
Disruptor Genie for 1min.
Verify cell disruption : place a 20mL under light microscope.
Disruptor Genie for 3min.
Verify cell disruption : place a 20mL under light microscope.
Centrifugation at 10,000 g; 30 min, 4°C. 
Removed the supernatant to clean 1.5mL Eppendorf tube, discard the pellet (cell debris). 
Centrifugation again at 10,000 g; 30 min, 4°C. 
Removed the supernatant to clean 1.5mL Eppendorf tube, discard the pellet (cell debris). 
Conserve the tubes at 4°C
