Team:Calgary/Notebook/Protocols/GlassBeadsCellLysisProtocolforProteinSamples

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<ul>
<li>0.1m diameter glass beads</li>
<li>0.1m diameter glass beads</li>
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<li>Lysis Buffer </li>
+
<li>Lysis buffer </li>
<li>Overnight induced cultures</li>
<li>Overnight induced cultures</li>
<li>1.5mL tubes</li>
<li>1.5mL tubes</li>

Revision as of 23:17, 27 September 2013

Glass Beads Cell Lysis Protocol for Protein Samples

Reagents and Materials

  • 0.1m diameter glass beads
  • Lysis buffer
  • Overnight induced cultures
  • 1.5mL tubes

Protocol

  1. Culture and induce protein-producing bacteria
  2. Centrifuge cells at max for 10 minutes. Discard supernatant
  3. Resuspend cells in 2mL of lysis buffer
  4. In a 1.5mL tube, add ~0.5mL of glass beads and 1mL of cells resuspended in lysis buffer
  5. Leave on ice for 5 min. Leave on BeadBeater or Vortex for 5 min. Repeat this cycle 6 times
  6. Centrifuge at max for 20 minutes
  7. Transfer supernatant (crude lysate) to a new tube and discard the glass beads

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