Patent ID: 7163809

Claim:
A method for producing a library of mutagenized polynucleotides from a target sequence, comprising: (a) taking a sample comprising (i) a target sequence including a section to be mutagenized, (ii) a library of first primers where the first primers include a first fixed sequence and a first unknown sequence 3′ to the first fixed sequence, the first unknown sequence varying within the library of first primers, and (iii) a library of second primers where the second primers include a fixed sequence that differs from the first fixed sequence; (b) performing one or more cycles of primer extension amplification on the sample in the presence of at least one polymerase such that a member of the library of the first primers is extended relative to the target sequence; and (c) performing one or more additional cycles of primer extension amplification on the sample such that a member of the library of the second primers is extended relative to the first primer that was extended in step (b) to form the library of mutagenized polynucleotides.