Patent ID: 8338091

Claim:
A method for preparing a recombinant nucleic acid molecule, the method comprising: (a) contacting two or more nucleic acid molecules, each nucleic acid molecule comprising two type IIs restriction enzyme recognition sites, with one or more type IIs restriction enzymes under conditions which allow for the generation of overhanging ends, to generate two or more digested nucleic acid molecules; (b) contacting a nucleic acid vector comprising two type IIs restriction enzyme recognition sites with one or more type IIs restriction enzyme under conditions which allow for the generation of overhanging ends and excision of a segment of the nucleic acid vector, to generate a digested nucleic acid vector, wherein the overhanging ends of the digested nucleic acid vector are each capable of hybridizing with at least one overhanging end of at least one of the two or more digested nucleic acid molecules; and (c) contacting the two or more digested nucleic acid molecules and the digested nucleic acid vector under conditions which allow for (i) hybridization of overhanging ends and (ii) covalent joining digested nucleic acid molecules to the digested nucleic acid molecule vector to form the recombinant nucleic acid molecule.