Materials
Materials
- Beckman Ultracentrifuge tubes (1 X 3.5 " or 9/16 " X 3 ½" )
- Beckman Ultracentrifuge fitted with a SW-28 or SW-41 rotor
- Serum Free DMEM
- PBS
- 10ml syringe
- 0.45 μm syringe filter tips
- 10% bleach
Methods
Methods
- Harvest virus with a 0.45um filter tip with a 10ml syringe, into a Beckman ultracentrifuge tube
- Bleach all virus producing cells, culture plates, syringes and filters with 10% bleach and allowed to stand for 45 minutes before discarding
- Balance all virus containing tubes with serum free DMEM (to 0.1g)
- Spin tubes at 25000rmp or 24000rmp for 90 mins in 4 degrees in a SW-28 or SW-41 rotor
- Discard supernatant into 10% bleach containing vessel
- Blot excess liquid with kimwipes. Turn tubes upside down in a rack and allow to drain for one minute
- Resuspend viral pellet in 100ul of PBS. Let virus sit overnight in 4 degrees
- Bleach all empty tubes with 10% bleach for45 minutes before discarding
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