Cell and Protein Extraction
- The solutions were centrifuged for 15 minutes at 4500 rpm at 4 degrees Celsius
- The pellet was collected and placed in a 50mL falcon tube
- The falcon tube was centrifuged at 4500 rpm for an additional 15 minute to precipitate the cells
- The black supernatant was removed (but not discarded) and placed into a 15mL falcon tube
- A small amount of 5mM Tris buffer was used to collect remaining traces of cells from the centrifuge tubes
- The pellet was then sonified
- 30 seconds signification followed by 30 seconds on ice until the pellet was loose and liquid
- Once liquefied, the black supernatant was put back in the tubes and the solution was centrifuged at 20,000rpm four 3 hours at 4C
- The supernatant was then placed in dialysis tubing
- The tube and its content were placed in 50mM Tris buffer in the cold room (50F) overnight to remove unwanted salts
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