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Methods for generating polynucleotides having desired characteristics by iterative selection and recombination

  • US 20030186356A1
  • Filed: 09/17/2002
  • Published: 10/02/2003
  • Est. Priority Date: 02/17/1994
  • Status: Abandoned Application
First Claim
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1. A method for introducing one or more mutations into a template double-stranded polynucleotide, wherein the template double-stranded polynucleotide has been cleaved into double-stranded random fragments of a desired size, comprising:

  • a) adding to the resultant population of double-stranded fragments one or more single or double-stranded oligonucleotides, wherein said oligonucleotides comprise an area of identity and an area of heterology to the template polynucleotide;

    b) denaturing the resultant mixture of double-stranded random fragments and oligonucleotides into single-stranded fragments;

    c) incubating the resultant population of single-stranded fragments with a polymerase under conditions which result in the annealing of said single-stranded fragments at regions of identity between the single-stranded fragments and formation of a mutagenized double-stranded polynucleotide; and

    d) repeating steps (b) and (c).

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