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Whole Bacteria Isolation from Kidney, Lung, or Spleen

Instrument: Bullet Blender or Bullet Blender Blue

Sample: Kidney, Lung, or Spleen tissue.

Protocol:

  1. Cut the organ tissue into appropriately sized pieces for analysis (50mg-300mg) and place into microcentrifuge tubes.
  2. Add a mass of 3.2 mm stainless steel beads equal to approximately 4x the mass of sample.  Each 3.2 mm stainless steel bead is approximately 140 mg.
  3. Add 300 μl of PBS into the tubes.
  4. Close the tubes. and place into the Bullet Blender®.
  5. Run at speed 8 for 4 minutes.
  6. Take out the tubes and inspect for complete homogenization of organ.  If homogenization is unsatisfactory, continue processing in two-minute increments until the sample contains no large chunks of tissue.
  7. Add an additional 200 μl of PBS to tube.  Run at speed 2 for 1 minute to vortex the samples.

Acknowledgement:  This protocol was developed by Hatice Karauezuem at Integrated Biotherapeutics Inc.