Ligation Protocol for Cloning with ElectroLigase® (M0369)

Protocol

1. Transfer ElectroLigase® and ElectroLigase® Reaction Buffer to ice prior to reaction set up. Mix tubes by finger flicking before use.
   
   
2. Combine 20–100 ng of vector* with a 3-fold molar excess of insert and adjust volume to 5 μl with dH₂O.
   
   
3. Add 5 μl of ElectroLigase Reaction Buffer and 1 μl of ElectroLigase and pipet up and down 7–10 times to mix.
   
   
4. Incubate ligation reaction at room temperature (25°C) for 30–60 minutes.
   
   
5. Chill sample on ice (if to be used within a few hours) or store at -20°C.
   
   * In-house testing has demonstrated that maximal transformation efficiency is achieved using between 20–100 ng of vector (blunt or sticky, including T-vectors) and a corresponding 3-fold molar excess of the insert to be ligated into the vector.