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• Homogenization of Cardiac Tissue

Homogenization of Cardiac Tissue

Instrument: Bullet Blender

Sample:  Cardiac Tissue

Protocol:

1. Cut heart into appropriately sized pieces for analysis (10mg-300mg). NOTE: Valves or blood vessels will require more vigorous homogenization due to their fibrous nature.
2. OPTIONAL: Wash tissue 3x with ~1mL PBS. Aspirate. NOTE: This step removes external contaminants (blood, etc.).
3. Sample
   1. Samples 50mg or greater. Place the sample in Navy bead lysis kit tube.
   2. Samples less than 50mg. Place the sample in Green bead lysis kit tube.
   3. Alternate protocol step for bulk beads. Place sample in microcentrifuge tube and add beads to the tube. Use a volume of beads equal to the mass of tissue. NOTE: 100mg ≅ 100μ
4. Add 0.025mL to 0.6mL buffer (2 volumes of buffer for every volume of sample).
5. Close the microcentrifuge tubes.
6. Place tubes into the Bullet Blender®.
7. Set controls for SPEED 8 and TIME 4 minutes. Press Start. (always make sure your tubes are balanced)
8. After the run, remove tubes from the instrument.
9. Visually inspect samples. If homogenization is unsatisfactory, run for another two minutes at SPEED 10.
10. Proceed with your downstream application.

Results: