Notes on the protocol: This protocol does not specify a particular buffer - you may choose which is most appropriate for your downstream application (nucleic acid isolation, protein extraction, etc.).
One of these Bullet Blenders
Bullet Blender 50-DX (BB50-DX)
Bullet Blender 50 Gold (BB50-AU)
2 x volume of sample
0.1 mm glass beads (GB01) Use a volume of beads equivalent to 1 x the volume of the sample
0.15 mm zirconium oxide beads (ZROB015) Use a volume of beads equivalent to 1 x the volume of the sample
If your sample has been grown on a plate or other solid surface, detach it (e.g, by flooding the plate with PBS and scraping) and place the material in a microcentrifuge tube. Liquid cultures may be placed directly in the tube as long as they are of sufficient density.
Centrifuge the suspension to yield a pellet. The pellet should be no larger than 3.5 ml.
Pipette off the supernatant, and resuspend the pellet in 2 volumes of homogenization buffer.
Place the sample in the tube with the beads.
Close the tubes tightly and place them in the Bullet Blender.
Set the controls for Speed 8 and Time 12. Press Start.
After the run, remove the tubes from the instrument and visually inspect the samples. If homogenization is incomplete, repeat the homogenization step at a higher speed.