Mammalian cell culture
Bullet Blender® homogenization protocol

  • Extract molecules (DNA, RNA, protein, chemicals)
  • Wet final product
  • Sample sizes: 100 to 3500 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.).

This protocol has been tested on brain tissue from a variety of animals. However, you may need to adjust speed/time settings for your particular sample due to the variation in tissue consistency from species to species.

Materials Required

uL

One of these Bullet Blenders

  • Bullet Blender 50-DX (BB50-DX)
  • Bullet Blender 50 Gold (BB50-AU)

Reagents

Homogenization buffer
2 x volume of sample

Bead choices

  • 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

Procedure

  1. 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.
  2. Centrifuge the suspension to yield a pellet. The pellet should be no larger than 3.5 ml.
  3. Pipette off the supernatant, and resuspend the pellet in 2 volumes of homogenization buffer.
  4. Place the sample in the tube with the beads.
  5. Close the tubes tightly and place them in the Bullet Blender.
  6. Set the controls for Speed 8 and Time 12. Press Start.
  7. 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.
  8. Proceed with your downstream application.