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Methods for the selection and cloning of nucleic acid molecules free of unwanted nucleotide sequence alterations

  • US 20030143605A1
  • Filed: 12/03/2002
  • Published: 07/31/2003
  • Est. Priority Date: 12/03/2001
  • Status: Abandoned Application
First Claim
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1. A method for eliminating an amplified nucleic acid molecule having a nucleotide sequence that is mutated, compared with the target nucleotide sequence used to produce the amplified nucleic acid molecule, comprising, (a) obtaining a mixture of duplexes of amplified nucleic acid molecules, wherein the duplex mixture comprises homoduplexes and heteroduplexes, (b) treating the duplex mixture with a single-strand specific nuclease that cleaves a mismatched site within a heteroduplex, and (c) subcloning the nuclease-treated duplex mixture, wherein a cleaved duplex is unsuitable for subcloning.

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