Patent ID: 7709626

Claim:
A method of making a fluorescently-labeled sequence-specific primer, comprising: adding a universal nucleic acid sequence to a sequence-specific primer, thereby generating a fluorescently-labeled sequence-specific primer, wherein the universal nucleic acid sequence comprises a 5′ end, a 3′ end; and a fluorescently-labeled nucleotide flanked on each side by at least one nucleotide, wherein nucleotides complementary to the nucleotides flanking the fluorescently-labeled nucleotide quench a detectable signal from the label fluorophore when the fluorescently-labeled sequence-specific primer is hybridized with its complementary nucleic acid molecule and the complementary nucleic acid is synthesized, wherein the sequence-specific primer can hybridize to a target nucleic acid sequence, wherein the fluorophore can be quenched by the nucleotides complementary to the nucleotides flanking the fluorescently-labeled nucleotide, and wherein the universal nucleic acid sequence does not substantially hybridize to the target nucleic acid sequence recognized by the sequence-specific primer.