Amplification of target nucleotide sequence without polymerase chain reaction
First Claim
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1. A method for detecting a nucleic acid comprising:
- hybridizing a first ligase chain reaction (LCR) primer and a second LCR primer to a target nucleotide sequence of a template nucleic acid such that the 3′
end of said first LCR primer is adjacent to the 5′
end of said second LCR primer;
ligating said first LCR primer to said second LCR primer to generate an LCR product;
hybridizing an RCA probe to at least a portion of said LCR product which includes the 3′
end of said LCR product;
initiating polymerization from said 3′
end of said LCR product, thereby generating an amplified single-stranded nucleic acid which comprises a plurality of LCR products; and
detecting said amplified single-stranded nucleic acid or fragments thereof.
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Abstract
The present disclosure relates to methods for amplifying nucleic acids and methods for detecting the amplified nucleic acid products. Nucleic acid amplification and detection can occur in solution or on a solid support.
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Citations
44 Claims
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1. A method for detecting a nucleic acid comprising:
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hybridizing a first ligase chain reaction (LCR) primer and a second LCR primer to a target nucleotide sequence of a template nucleic acid such that the 3′
end of said first LCR primer is adjacent to the 5′
end of said second LCR primer;
ligating said first LCR primer to said second LCR primer to generate an LCR product;
hybridizing an RCA probe to at least a portion of said LCR product which includes the 3′
end of said LCR product;
initiating polymerization from said 3′
end of said LCR product, thereby generating an amplified single-stranded nucleic acid which comprises a plurality of LCR products; and
detecting said amplified single-stranded nucleic acid or fragments thereof. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A kit comprising:
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a chip having a capture probe attached thereto;
a detector reagent; and
instructions for using said detector reagent to detect an amplified target nucleotide sequence attached to said chip via said capture probe. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
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Specification