Homogenization of Drosophila Larvae
Instrument: Bullet Blender or Bullet Blender Blue
Sample: Fruit fly (D. melanogaster) larvae
- If you have not already, wash Drosophila larvae 3x with 1ml PBS or other buffer, as appropriate, to remove food, surface bacteria, and other contaminants.
- Aspirate the larvae, or remove as much liquid as possible with a pipette.
- Place 10 - 300 mg of Drosophila larvae into microcentrifuge.
- Add a volume of 0.5mm glass beads (zirconium silicate beads are an appropriate substitute for DNA applications) equal to the mass of tissue. A useful guideline is 100mg ≅ 100μL, as the flies may lie loose in the tube and occupy more volume than they would compressed.
- Add 2 volumes of buffer for every volume of sample.
- Securely close the microcentrifuge tubes closed and place them into the Bullet Blender®.
- Set the Bullet Blender to a speed of 8 and a run time of 3 minutes. Press start.
- After the run, remove the tubes from the instrument.
- Visually inspect samples, if homogenization is unsatisfactory, run for another two minutes at speed 10.