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Methods for purifying annealed double-stranded oligonucleotides lacking base pair mismatches or nucleotide gaps

  • US 6,673,552 B2
  • Filed: 02/14/2002
  • Issued: 01/06/2004
  • Est. Priority Date: 01/14/2002
  • Status: Expired due to Term
First Claim
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1. A method for purifying double-stranded polynucleotides lacking base pair mismatches, insertion/deletion loops and/or a nucleotide gap or gaps comprising the following steps:

  • (a) providing a plurality of polypeptides that specifically bind to a base pair mismatch, an insertion/deletion loop and/or a nucleotide gap or gaps within a double stranded polynucleotide comprising a Vsr (very short patch repair) bacterial DNA mismatch repair enzyme;

    (b) providing a sample comprising a plurality of double-stranded polynucleotides;

    (c) contacting the double-stranded polynucleotides of step (b) with the polypeptides of step (a) under conditions wherein a polypeptide of step (a) can specifically bind to a base pair mismatch, an insertion/deletion loop and/or a nucleotide gap or gaps in a double-stranded polynucleotide of step (b); and

    (d) separating the double-stranded polynucleotides lacking a specifically bound polypeptide of step (a) from the double-stranded polynucleotides to which a polypeptide of step (a) has specifically bound, thereby purifying double-stranded polynucleotides lacking base pair mismatches, insertion/deletion loops and/or a nucleotide gap or gaps.

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