Patent ID: 7575860

Claim:
A single step method of joining two or more linear duplex DNA molecules comprising incubating two or more linear duplex DNA molecules, each having an x and x′ strand of opposite polarities, wherein the 5′ end of the x′ strand of one linear duplex DNA molecule comprises a sequence of nucleotides that are complementary between about 5 and about 100 nucleotides to the 5′ end of the x strand of the linear duplex DNA molecule to which it is to be joined, wherein said incubation is in the presence of a DNA polymerase under conditions whereby the two or more linear duplex DNA molecules are joined, the DNA polymerase has intrinsic 3′-5′ exonuclease activity and joins the DNA molecules in an exonuclease-mediated reaction that is mediated by said DNA polymerase and said two or more linear duplex DNA molecules have 5′-overhanging, 3′-overhanging and/or blunt ends.