7.2.2. High speed centrifugation
1. Centrifuge the supernatant
at 75,000 g and 4oC for 3
2. Discard the supernatant.
3. Re-suspend the pellet 5 ml extraction buffer.
This is best achieved by storing the pellet with buffer overnight at 4oC, to loosen the pellet.
4. Next day vortex the pellet lightly.
5. Centrifuge at 8,000 g and 4oC for 15 minutes.
6. Retain the supernatant for gradient centrifugation.