Amplification method for a polynucleotide
First Claim
1. A method for amplification of at least a portion of a target nucleic acid comprised of a first strand, comprising(a) at a first location, generating based on the first strand copies of a second strand complementary to the first strand, which copies of the second strand are, or are adapted to be, captured on a solid support,(b) moving the copies of the second strand to a second location either captured on the solid support or such that said copies of the second strand are captured on the solid support at the second location, and(c) at the second location, generating based on the copies of the second strand copies of at least a portion of the first strand.
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Accused Products
Abstract
The nucleic acid amplification procedures of the present invention provide methods for amplification of a nucleic acid comprised of a first strand, comprising (a) using the first strand to generate copies of a second strand at a first location, (b) moving the copies of the second strand to a second location, and (c) using the copies of the second strand to generate copies of at least a portion of the first strand. Target nucleic acids used in the context of the present method include RNA or DNA, either single stranded or double stranded, using primer extension or joining-type protocols. Embodiments are set forth for automated forms of the claimed procedures.
175 Citations
30 Claims
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1. A method for amplification of at least a portion of a target nucleic acid comprised of a first strand, comprising
(a) at a first location, generating based on the first strand copies of a second strand complementary to the first strand, which copies of the second strand are, or are adapted to be, captured on a solid support, (b) moving the copies of the second strand to a second location either captured on the solid support or such that said copies of the second strand are captured on the solid support at the second location, and (c) at the second location, generating based on the copies of the second strand copies of at least a portion of the first strand.
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25. A method for amplifying one or more segments of a target nucleic acid, wherein the nucleic acid comprises a sequence that is complementary to a first and a second probe, the method comprising:
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(a) providing the target nucleic acid attached at a first location; (b) adding first, second, third, and fourth probes to the target nucleic acid under conditions that allow hybridization between complementary strands of nucleic acid, and such hybridization occurs, wherein the 3'"'"' end of one and the 5'"'"' end of the other of the first and second probes or the 3'"'"' end of one and the 5'"'"' end of the other of the third and fourth probes are adjacent to each other upon hybridization of the first and second probes or third and fourth probes to the target nucleic acid, wherein the first and second probes are complementary to a polynucleotide consisting of the third and fourth probes joined together, and wherein the second and fourth probes include a first moiety and a second moiety, respectively; (c) ligating or chemically joining those of the first and second probes or third and fourth probes which are made adjacent by hybridizing to the target nucleic acid; (d) washing unhybridized probes from the first location; (e) denaturing to separate the joined probes from the target nucleic acid; (f) moving the denatured ligated first and second probes or denatured ligated third and fourth probes from the first location to a second location; and (g) generating at the second location the complement of a least of portion of said moved, ligated probes. - View Dependent Claims (26, 27, 28, 29)
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30. A method for amplifying at least a portion of a target nucleic acid, wherein the nucleic acid comprises a sequence that is complementary to a first and a second probe, the method comprising:
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(a) attaching a target nucleic acid at a first location; (b) combining first and second probes with the target nucleic acid under conditions that allow hybridization between complementary strands of nucleic acid, and hybridizing the first and second probes to the target nucleic acid so that the 3'"'"' end of one and the 5'"'"' end of the other of the first and second probes are adjacent to each other upon such hybridization, wherein the second probe includes a first moiety; (c) ligating or chemically joining the adjacent first and second probes; (d) washing unhybridized probes from the first location; (e) denaturing to separate the joined probes from the target nucleic acid; (f) moving the denatured joined first and second probes from the first location to a second location; and (g) generating at the second location the complement of a least of portion of said moved, ligated probes.
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Specification