Patent ID: 7052876
Filing Date: 2006-05-30
Classification: C07K,C12N

Abstract:
1. A method of generating a library of DNA molecules of varying length and sequence in a desired orientation comprising the steps of: a) providing a mixture of double-stranded DNA molecules, each of said molecules having 5′ and 3′ ends which are compatible to each other and are equivalent to the cleavage products of different but compatible restriction enzymes; and b) allowing ligation to take place, wherein ligation of said double-stranded DNA molecules in desired orientations generates molecules that are not cut by either of said restriction enzymes whereas ligation in undesired orientations generates molecules that retain at one or more ligation points a restriction site that is recognised by one of said restriction enzymes; and c) cutting the ligated DNA molecules with one or both of said restriction enzymes such that only molecules that are ligated in undesired orientations are cut, leaving a library of DNA molecules of varying length and sequence in a desired orientation.