Patent ID: 7838219

Claim:
An in vitro method of generating mutations in a zone around a particular nucleotide within a desired polynucleotide comprising: (a) preparing at least one partially complementary polynucleotide having at least one non-complementary nucleotide within the corresponding zone of the desired polynucleotide; (b) forming at least one heteroduplex between said desired polynucleotide and said partially complementary polynucleotide; (c) combining said heteroduplex with at least one nucleotide analogue and an effective amount of an agent or agents with exonuclease activity, polymerase activity, and mismatch-directed strand cleavage activity, wherein said mismatch-directed strand cleavage activity is provided by the protein of SEQ ID NO: 34 (RES I); (d) allowing sufficient time for the nucleotide analogue to be incorporated into either said desired polynucleotide, said partially complementary polynucleotide, or both polynucleotides; and (e) replicating a polynucleotide of step d such that mutations are generated in a zone around a particular nucleotide within the desired polynucleotide.