Why are 2 different restriction enzymes used when preparing genome libraries?

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Michael J. Benedik

You don't need to use 2 different enzymes. Many (perhaps most) genomic libraries are made with a single enzyme. You want to use 2 enzymes when you need directional cloning.

Ajay R. Bhat

Digestion with 2 restriction enzymes will avoid the self ligation or re-circularization of the plasmid/vector. If you are using single enzyme, you can use alkaline phosphatase to avoid re-circularisation.