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• Honeybee Thorax Homogenization in the Bullet Blender

Honeybee Thorax Homogenization in the Bullet Blender

Instrument: Bullet Blender or Bullet Blender Blue

Sample: Honeybee (Apis mellifera) thoraces

Protocol:

1. If you have not already, wash the A. mellifera 3x with PBS or other buffer, as appropriate, to remove surface contaminants.
2. Isolate the thorax using a scalpel and forceps
3. Place each thorax, individually, into a microcentrifuge tube.
4. Add 100 mg of 1.0 mm glass beads (approx. 40 µl) to each microcentrifuge tube.
5. Add 600 µl of lysis buffer (100 mM Tris, pH 8.0,10 mM EDTA, pH 8.0, and 1% SDS) to each tube.
6. Securely close the microcentrifuge tubes closed and place them into the Bullet Blender®.
7. Set the Bullet Blender to a speed of 8 and a run time of 3 minutes. Press start.
8. After the run, remove the tubes from the instrument.
9. Visually inspect samples, if homogenization is unsatisfactory, run for another two minutes at speed 10.

Refeerence:

Bourgeois, A.L., Rinderer, T.E. Genetic Characterization of Russian Honey Bee Stock Selected for Improved Resistance to Varroa destructor. J. Econ. Entomol. 102(3):1233-1238. 2009.