Polynucleotide and use thereof
AssigneeSamsung Electronics Co., Ltd.
InventorsSea-hee Kim, Joon-ho Kim, Joo-won Rhee, Sun-ok Jung, Ko-bong Choi, Kyu-youn Hwang
Life of this patent
Estimating the term…
Abstract
A polynucleotide comprising a first region the 5′ end of which is complementary to a portion of a target nucleic acid, a cleavable second region, a third region having a stem-loop structure, and a fourth region complementary to the 3′ end of the first region, and use of the polynucleotide, as well as a composition comprising two such polynucleotides each of which hybridize different strands of a double-stranded target nucleic acid, and methods and kits using the same for amplifying targets.
Claims14
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.
Dependencies are inferred from the claim text.