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US 6,582,914 B1Granted
Method for generating a library of oligonucleotides comprising a controlled distribution of mutations
AssigneeGenencor International, Inc.
InventorsRobert M. Caldwell, Volker Schellenberger
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Abstract
Methods are disclosed for producing libraries of nucleic acid molecules which libraries are derived from a nucleic acid template. The libraries comprise variant nucleic acids which are produced from a mutagenesis strategy using, e.g., a plurality of defined mutagenic and/or non-mutagenic primers and specific reaction conditions which favor the production of varied combinatorial mutants.
Claims10
A method of producing a library of mutant nucleic acid molecules comprising:
- (a) obtaining a template nucleic acid;
- (b) preparing an oligonucleotide primer pair corresponding to the ends of said template nucleic acid;
- (c) preparing a first and a second mutagenic oligonucleotide primer, wherein said first primer corresponds to a first desired mutation within said template nucleic acid, and said second primer corresponds to a second desired mutation within said template nucleic acid;
- (d) mixing the oligonucleotide primers prepared in said steps (b) and (c);
- and
- (e) combining said mixture in said step (d) with the template nucleic acid under conditions to facilitate the polymerase chain reaction, wherein said mutagenic oligonucleotides are present in a concentration that is less than saturation concentration.
3 more elements
Dependencies are inferred from the claim text.