What are the best conditions for DNA ligation?

Find out how the downstream application dictates the best reaction conditions for ligation.


Eric Cantor:

Optimal reaction conditions depend on the downstream step. For library construction, low temperatures such as 16 degrees Celsius and long incubation times, 12-16 hours, provide the best reaction yields. This allows you to maximize library complexity. Complexity is not important for subcloning of a single insert into a vector; therefore, you can accomplish this with a much shorter reaction time, 15 minutes or less, over a much broader range of temperatures, 16 degrees Celsius to 37 degrees Celsius.

Related Videos

  • DNA Ligase
    DNA Ligation
  • BestRatios_thumb
    What molar ratios should I use for DNA Ligation?
  • WhyUsePEG_thumb
    Why do I need to add PEG to my DNA ligation?
Visit NEB’s Video Library
Loading Spinner