Oneill Lab:Enzymes

Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology

Lab Stock Enzymes

Preparation of lab stock enzymes and buffers

10x EcoRI Buffer

1. Combine
   • 8mL 2.5M Tris
   • 2mL 5M NaCl
   • 1mL 1M MgCl₂
   • 50μL TritonX-100
2. Adjust pH to 7.5 with 12M HCl
3. Add water to 20mL

Proteinase K

Makes 0.5mL at 20mg/ml

• 10μL 1M CaCl₂
• 5μL 1M Tris pH 7.5
• 250μL Glycerol
  • Pipette with cut tip
  • Use under flame
• 10mg ProK
  • Stored in 4°C dessicator
• 235μL H₂O

Aliquot and store at -20°C