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

Nuclease Solution

Used in lambda extractions prior to capsid degradation.

• DNaseI 10mg
• RNaseA 10mg
• 1M Tris 10μl
• 5M NaCl 30μl
• 1M MgCl₂ 1μl
• Water 950μl

Use at concentration of 1mg/ml

Links and Contacts

Seth Kasowitz [1]

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