Patent ID: 6159687
Filing Date: 2000-12-12
Classification: C12N

Abstract:
A method of shuffling polynucleotides, comprising(a) providing an isolated sense strand from at least a first variant form of a polynucleotide, and an isolated antisense strand from at least a second variant form of the polynucleotide, wherein the isolated sense and antisense strands are starting substrates to be shuffled; and(b) shuffling the isolated sense and antisense strands provided in step (a) by contacting the strands with at least one primer or one pair of primers and conducting a multi-cyclic polynucleotide extension reaction wherein(i) in at least one cycle, the primer(s) anneal to the isolated strands and generate extended primers; and(ii) in at least one subsequent cycle, the extended primers generated in a previous cycle act as both primers and templates to form further extended primers,whereby the polynucleotide extension is continued for sufficient cycles until the further extended primers includes recombinant forms of the shuffled polynucleotides.