E. coli DNA Ligase Protocol (M0205)

1. Set-up the reaction as follows:
DNA up to 5 µg
E. coli DNA Ligase Reaction Buffer (10X)
2 µl
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.
  1. NEBioCalculator Ligations Thumbnail

    NEBioCalculator® - Using the Ligation module

    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.