Patent ID: 9476078
Filing Date: 2016-10-25
CPC Classification: C12N,C12P

Claim Text:
1. A method of producing a plurality of modified polynucleotides having more than one mutation per polynucleotide, comprising: (a) adding at least three primers to a double stranded template polynucleotide in a single reaction mixture, wherein the at least three primers are not overlapping, and wherein each of the at least three primers comprise at least one mutation different from the other primers, wherein at least one primer is a forward primer that can anneal to a minus strand of the template and at least one primer is a reverse primer that can anneal to a plus strand of the template, and (b) subjecting the reaction mixture to a polymerase extension reaction to yield a plurality of extended modified polynucleotides having more than one mutation per polynucleotide from the at least three primers; wherein the modified polynucleotides comprise all possible combinations of mutations to the template polynucleotide.