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Homogenization of C. elegans

Instrument: Bullet Blender® STORM 5

Sample: Caenorhabditis elegans.  May be larval, dauer, or adult.

Protocol:

1. Harvest worms from culture plate by washing (either with saline or water) into a 5 ml tube.
2. Centrifuge worm suspension at 200-500g for five minutes to yield a pellet under the washing liquid.
3. Completely aspirate the supernatant liquid.
4. Inspect the volume of the pellet. It should be 0.75 ml 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 speed to 8 and 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.