IGEM:UC Berkeley/2006/Ligation

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. Create Ligation mixture:
   • 69uL H2O
   • 10uL Ligase buffer
   • 10 uL backbone
   • 10uL insert
   • 1uL Ligase
2. Incubate at room temperature for 60 minutes.