Jonesma Lab:Protocols:ligation

1. Thaw reagents and store on ice

2. Combine the following reagents

      2 μL Ligation Buffer 

      5 μL Digested backbone

     11 μL Digested insert (smaller piece)

      2 μL T4 DNA ligase

3. Incubate at 22°C for 1 hr, 65°C for 10min (Use a PCR machine for this)

4. Transform bacteria with your ligation