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Analysis of nucleotide polymorphisms at a site

  • US 6,762,018 B1
  • Filed: 12/23/1999
  • Issued: 07/13/2004
  • Est. Priority Date: 12/23/1999
  • Status: Expired due to Fees
First Claim
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1. A method for determining the identity of a polymorphic nucleotide in a target sequence having at least two known variant nucleotides at a site, comprising:

  • performing a primer extension reaction with the target sequence using an extension reaction mixture comprising;

    a primer that specifically hybridizes to the target sequence such that there is a one or more nucleotide gap between the 3′

    terminus of the primer and the variant nucleotide of the polymorphic site of the target sequence, and a mixture of deoxyribonucleoside triphosphates (dNTPs) or ribonucleoside triphosphates (rNTPs), where the mixture of dNTPs or rNTPs provide for at least one nucleotide extension of the primer when hybridized to a target sequence having either of the two variant nucleotides at the polymorphic site, wherein the mixture excludes a dNTP or rNTP complementary to one of said variant nucleotides of the polymorphic site, and wherein the dNTPs or rNTPs in the mixture are not detectably labeled or modified, and wherein the extension reaction is performed in the absence of a dideoxynucleoside triphosphate (ddNTP); and

    analyzing primer extension products of said extension reaction;

    wherein the length of the primer extension products is indicative of the identity of the variant nucleotides at the polymorphic site.

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