How does reaction time effect ligation?

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Yuan-Yeu Yau

Hi Gregory,

For sticky-end ligation, I usually use 2-3 hours at room temperature. But, sometimes, if I am not in a hurry, I will do overnight. For blunt-end ligation, most of the time I will do overnight. To my experience, I will not consider 1hr or 24hr ligation really critical. There are other critical things to concern in ligation.Ligation experiments are either successful or fail. You can have a plate with 1,000 colonies or with 10 colonies. But, with one positive clone, then you are done.

Gregory James Buchan

Thank you! I have noticed that to be the case and I just wanted to make sure!

You are welcome. I saw your another question 'disappearing plasmid'. Have you got your plasmid yet from the miniprep sample since? I just curious. I also sent you a couple of suggestions reguarding the question.