Detection of nucleic acid variation by cleavage-amplification (CleavAmp) method
First Claim
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1. A method for detecting a polymorphism in a polynucleotide, comprising:
- (a) annealing a probe to a region of a polynucleotide suspected of containing a polymorphism to form a complex, wherein the probe comprises a non-extendable 3′
end and is not complementary to the polymorphism;
(b) contacting the complex with an enzyme or chemical that cleaves the probe and the polynucleotide at a region of mismatch between the probe and the polynucleotide to produce a probe with an extendible 3′
end;
(c) adding an artificial template, wherein the cleaved probe acts as a primer for amplifying the artificial template; and
(d) amplifying the artificial template, wherein the presence of an amplified product indicates the presence of the polymorphism.
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Abstract
Methods and compositions for detecting nucleic acid polymorphisms are provided.
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Citations
27 Claims
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1. A method for detecting a polymorphism in a polynucleotide, comprising:
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(a) annealing a probe to a region of a polynucleotide suspected of containing a polymorphism to form a complex, wherein the probe comprises a non-extendable 3′
end and is not complementary to the polymorphism;
(b) contacting the complex with an enzyme or chemical that cleaves the probe and the polynucleotide at a region of mismatch between the probe and the polynucleotide to produce a probe with an extendible 3′
end;
(c) adding an artificial template, wherein the cleaved probe acts as a primer for amplifying the artificial template; and
(d) amplifying the artificial template, wherein the presence of an amplified product indicates the presence of the polymorphism. - 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 method for detecting nucleotide variations between target nucleic acid comprising:
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(a) preparing a gene specific probe with an non-extendable 3′
end, wherein the probe is complementary to a region of the target nucleic acid;
(b) hybridizing the gene specific probe to the target nucleic acid to form a duplex, wherein a variation structure is formed in the duplex if the target nucleic acid comprises a nucleotide variation;
(c) exposing the duplex to a cleavage enzyme or chemicals, wherein the enzyme or the chemicals cleave the variation structure in the duplex to remove the non-extendable 3′
end from the gene specific probe and generated a new extendable 3′
end on the probe and the target nucleic acid; and
(d) amplifying an artificial template using the cleaved gene specific probe or target nucleic acid as primers. - View Dependent Claims (27)
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Specification