Use of codon-varied oligonucleotide synthesis for synthetic shuffling
AssigneeMaxygen, Inc.
InventorsMark Welch, Jon Ness, Claes Gustafsson, Willem P. C. Stemmer, Jeremy Minshull
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Abstract
Methods of providing shuffling libraries that include codon-varied oligonucleotide sequences are described. Codon-varied oligonucleotides are synthesized using trinucleotide or mononucleotide phosphoramidite sequences, and are derived from homologous or non-homologous nucleic acid sequences, or combinations of such sequences. Various methods of recombining codon-varied oligonucleotide sequences to expedite artificial evolution are also described. The present invention additionally relates to various integrated systems that are optionally used to automate these recombination methods.
Claims39
A method of providing a population of recombined nucleic acids, the method comprising:
- (i) providing a population of oligonucleotides comprising at least one set of codon-varied oligonucleotides, wherein at least one member of the set of codon-varied oligonucleotides is chemically synthesized using at least trinucleotide sequences and wherein two or more members of the population comprise overlapping oligonucleotides;
- (ii) hybridizing at least two of the overlapping oligonucleotides to each other to provide a population of hybridized overlapping oligonucleotides, which population of hybridized overlapping oligonucleotides comprises at least one codon-varied oligonucleotide;
- and,
- (iii) elongating members of the population of hybridized overlapping oligonucleotides, thereby providing the population of recombined nucleic acids.
Dependencies are inferred from the claim text.