Jonesma Lab:Protocols:ligation

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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