E. coli DNA Ligase Protocol (M0205)
|DNA||up to 5 µg|
|E. coli DNA Ligase Reaction Buffer (10X)
|E. coli DNA Ligase||1 µl (10 units)|
|H2O||up to 20 µl|
2. Incubate at 16°C for 30 minutes.
3. Heat inactivate by incubating at 65°C for 20 minutes.
This tutorial describes the use of the NEBioCalculator web tool to optimize the molar ratio between vector and insert DNA for use in a ligation reaction.