Oneill Lab:Enzymes

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

Contents

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]

Lab Homepage

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