Patent ID: 9428798
Filing Date: 2016-08-30
CPC Classification: C12Q

Claim Text:
1. A method of amplifying a target nucleic acid, the method comprising: incubating or heating a first polynucleotide and a second polynucleotide in the presence of an endonuclease to provide a first incubated or heated reactant, wherein the first polynucleotide comprises: a first region comprising at least two consecutive nucleotides including the 5′-terminus that are complementary to at least two consecutive nucleotides of a first strand of a double-stranded target nucleic acid sequence; a second region located 3′ of the first region comprising at least one cleavable nucleotide sequence, a third region located 3′ of the second region that comprises a nucleotide sequence with a stem-loop structure, and a fourth region located 3′ of the third region comprising at least two consecutive nucleotides complementary to at least two consecutive nucleotides of the 3′ end of the first region, wherein the second polynucleotide comprises a first region comprising at least two consecutive nucleotides including the 5′-terminus of the second polynucleotide that are complementary to at least two consecutive nucleotides of a second strand of the double stranded target nucleic acid sequence, a second region located 3′ of the first region comprising at least one cleavable nucleotide sequence, a third region located 3′ of the second region that comprises a nucleotide sequence with a stem-loop structure, and a fourth region located 3′ of the third region comprising at least two consecutive nucleotides of the 3′ end of the first region, wherein the incubation or heating of the first polynucleotide and the second polynucleotide in the presence of the endonuclease nicks the second regions of the first polynucleotide and the second polynucleotide; incubating or heating the first incubated or first heated reactant to form a single-stranded first region of the first polynucleotide and a single-stranded first region of the second polynucleotide; incubating the single-stranded first region of the first polynucleotide, the single-stranded first region of the second polynucleotide, and a double stranded target nucleic acid to prepare a hybridized product comprising the first region of the first polynucleotide hybridized to the first strand of the target nucleic acid, and the first region of the second polynucleotide hybridized to the second strand of the target nucleic acid; and incubating the hybridized product in the presence of a nucleic acid polymerase to amplify the target nucleic acid.