Organs (for bacterial extraction)
Bullet Blender® homogenization protocol

  • Release bacteria, viruses, organelles
  • Wet final product
  • Sample sizes: 50 to 300 mg.

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.).

Materials Required

uL

One of these Bullet Blenders

  • Bullet Blender (BBX24)
  • Bullet Blender Blue (BBX24B)
  • Bullet Blender Storm 24 (BBY24M)
  • Bullet Blender 24 Gold (BB24-AU)

Reagents

Homogenization buffer
2 x volume of sample
PBS (optional)
2 x volume of sample

Bead choices

  • 3.2 mm stainless steel beads (SSB32) Use a volume of beads equivalent to 1 x the volume of the sample

Procedure

  1. Cut the sample into appropriately sized pieces. For larger samples, we recommend cutting the material into long, thin strips for faster homogenization.
  2. Wash the sample 3x with 1/2 tube volume of PBS to remove surface contaminants.
  3. Place the sample in the tube with the beads.
  4. Add a volume of buffer that is twice the volume of the sample. Sample volume may be approximated by sample weight. E.g., for a 100 mg. sample, add 0.2 ml. buffer.
  5. Close the tubes tightly and place them in the Bullet Blender.
  6. Set the controls for Speed 8 and Time 3. Press Start.
  7. Set the controls for Speed 8 and Time 5. Press Start.
  8. Add an additional 200 ul of buffer to the tubes, and return them to the Bullet Blender.
  9. Set the controls for Speed 3 and Time 1. Press Start.
  10. Proceed with your downstream application.