Team:Calgary/Notebook/Protocols/GlassBeadsCellLysisProtocolforProteinSamples

From 2013.igem.org

Revision as of 01:22, 29 October 2013 by Cmwinter (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Glass Beads Cell Lysis Protocol for Protein Samples

Reagents and Materials

  • 0.1mm 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 1:10 dilution of lysis buffer (Lysis buffer composed of: 50mM NaH2PO4, 300mM NaCl, 10mM imidazole, pH 8.0).
  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 2 times.
  6. 6. Centrifuge at 14,000rpm for 20 minutes.
  7. Transfer supernatant (crude lysate) to a new tube and discard the glass beads.