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.