Patent ID: 7588896

Claim:
A method of determining a target polynucleotide comprising; providing a target polynucleotide, a primer set, and a hairpin linker, wherein the primer set comprises a forward primer and a reverse primer, wherein the reverse primer comprises a target specific portion, a single-stranded stem portion, and a protruding tail, wherein the forward primer comprises a target specific portion, and a single-stranded stem portion, wherein the hairpin linker comprises a loop portion, a stem portion, and a protruding tail, wherein the single-stranded stem portion of the forward primer is complementary to the complement of the single-stranded stem portion of the reverse primer, wherein the protruding tail of the hairpin linker is complementary to the protruding tail of the reverse primer; amplifying the target polynucleotide in a polymerase chain reaction (PCR) using the forward primer and the reverse primer to form a first amplification product; melting the first amplification product; hybridizing the single-stranded stem portion of the forward primer to the complement of the single-stranded stem portion of the second primer to form a self-complementary amplification product; hybridizing the protruding tail of the self-complementary amplification product to the protruding tail of the hairpin linker; ligating the self-complementary amplification product to the hairpin linker to form a loop ligation product; detecting the loop ligation product; and, determining the target polynucleotide.