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Abstract

The present invention provides methods and kits for determining the identity of a nucleotide at a variant site in a target nucleic acid of interest, including, for example, point mutations and single nucleotide polymorphisms. The methods involve conducting template-dependent extension reactions in the presence of a mixture of nucleotides that include at least one labeled extendible nucleotide and at least one labeled non-extendible nucleotide that are selected to be complementary to the nucleotides that potentially occupy the variant site. The particular labeled nucleotide incorporated into the extension products is characteristic of the nucleotide at the variant site. In addition to their utility in analyzing point mutations and single nucleotide polymorphisms, the methods and kits of the invention have utility in a variety of other applications in which specific nucleotide sequence information is of value, including, for example, paternity disputes, prenatal testing, forensic analysis and detection of pathogens.

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First Claim