Patent ID: 7078168

Claim:
A method of determining the frequency of an allele in a population of nucleic acid molecules, said method comprising: pooling the nucleic acid molecules of said population to provide a pooled nucleic acid sample; performing primer extension reactions in a reaction mixture comprising said pooled nucleic acid sample and a primer which binds at a predetermined site located in said nucleic acid molecules, wherein said site is substantially adjacent to a polymorphic position of interest in said allele, to provide primer extension products, and wherein the primer extension reaction is performed by sequentially adding non-chain terminating nucleotides to the reaction mixture and quantitatively determining the incorporation or non-incorporation of each nucleotide as each nucleotide is added by bioluminometrically detecting the release of pyrophosphate; obtaining a pattern of nucleotide incorporation in said primer extension products at the positions that correspond to said polymorphic position of interest; and determining the frequency of said allele from said pattern of nucleotide incorporation.