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Homogenization of Pine Nut

Instrument: Bullet Blender

Sample:  Pine Nut

Protocol:

  1. Place one nut (100-200mg) into a microcentrifuge tube.
  2. OPTIONAL: Wash nut 3x with ~1mL PBS. Aspirate. NOTE: This step removes some external contaminants and debris.
  3. Add 0.6g of zirconium oxide beads (0.5mm) to the tube.
  4. Add 0.4mL buffer to the tube.
  5. Close the microcentrifuge tubes.
  6. Place tubes into the Bullet Blender®.
  7. Set controls for SPEED 10 and TIME 4 minutes. Press Start.
  8. After the run, remove tubes from the instrument.
  9. Visual inspection of the homogenate is difficult due to the formation of emulsions with the oils from the plant tissue in aqueous buffers. To verify sufficient homogenization, you may place forceps or a spatula inside the tube to search for remaining large fragments. If homogenization is unsatisfactory, run for another two minutes at the SPEED 10.
  10. Remove sample tubes from the Bullet Blender® and proceed with your downstream application.

Results: