Patent ID: 8129119

Claim:
A method for introducing sequence changes into a nucleic acid molecule, the method comprising steps of: contacting a template nucleic acid molecule with a first primer containing a first portion that hybridizes with the template nucleic acid molecule, a second portion that includes at least one terminating residue that does not serve as a template for at least one polymerase, wherein the terminating residue comprises a 2′-O-methyl residue, and a third portion that contains the sequence changes as compared with the template nucleic acid molecule complement; extending the first primer to produce a first modified strand; and contacting the first modified strand with a second primer that hybridizes thereto; extending the second primer with the polymerase, so that a double-stranded product molecule is generated containing a 5′ overhang due to failure of the terminating residue to serve as a template for the polymerase.