IGEM:UC Berkeley/2006/Ligation

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Basic Ligation

  1. Create Ligation Mixture:
    • 5.5 uL water
    • 1 uL ligation buffer
    • 0.5 uL ligase
    • 1 uL backbone (pAC997)
    • 2 uL insert
  2. Let ligate at room temp for 30 minutes
    • Note: For the control, add 2 uL of water instead of the insert

Scaled-Up Ligations:

  • Use this for the construction of libraries.
  1. 69uL H2O
  2. 10uL Ligase buffer
  3. 10 uL backbone
  4. 10uL insert
  5. 1uL Ligase
  6. Incubate at room temperature for 30 minutes.
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