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Homogenization of Saccharomyces Cultures

Instrument: Bullet Blender® STORM 5

Sample: Budding yeast (S. cerevisiae, S. pombe)

Protocol:

1. Pour overnight yeast culture into a 5ML tube.
2. Centrifuge culture to yield a cell pellet (1000g for two minutes).
3. Completely aspirate the supernatant liquid. Place tube on ice.
4. Inspect the volume of the pellet. It should be 0.75ML or less in order to achieve efficient homogenization.
5. Add a volume of 0.5mm zirconium oxide beads equal to the volume of cells.
6. Add 2 volumes of buffer for every volume of sample.
7. Tightly screw the centrifuge tubes closed and place them into the Bullet Blender®.
8. Set the speed to 8 and the time to 3 minutes. Press start.
9. After the run, remove the tubes from the instrument.
10. Visually inspect samples.  If homogenization is unsatisfactory, run for another two minutes at speed 9.