Question

Early gene cloning experiments involved insertion at one restriction site in the vector; for example, the insert would have and EcoRI site at each end, and he vector would be opened at an RI site prior to litigation. Under what circumstances would asymmetric cloning be desirable, with the insert having a different site at each end?

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Answer 1

Asymmetrical cloning where the insert has a different site at each end is desirable when it is to be cloned in a vector and inserted between two different restriction sites.

For example, the vector has a BamH1 and EcoR1 site and the insert is to be added between these two, so we treat the insert with these two restriction enzymes so it has a bAmh! at one end and EcoR1 site at the other end, so that it can be easily ligated to the vector.