When you ligate an insert into one restriction site of a vec

When you ligate an insert into one restriction site of a vector, what would you do to reduce self-ligation of the vecor? Explainwhy.

Solution

Unless you are using a blunt end cloning strategy or digestion with a single restriction enzyme, self-ligations should not be a problem. There are also excellent ligation-independent, recombination-based cloning strategies (Gibson cloning, Gateway cloning) that eliminate the problems of recovering empty vector.Use phosphatase treatment if its involving Coheshive/ Sticky ends.Using a gradient of Vector : Insert ratio(1:3 or 1:6), may yield less self ligation.