Endonuclease-enhanced helicase-dependent amplification
First Claim
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1. A method for assaying a sample, the method comprising combining a composition comprising a restriction endonuclease, a helicase, a DNA polymerase, and at least a first primer and a second primer,with a sample containing or potentially containing a nucleic acid that hybridizes to the first and second primer, andamplifying the target sequence to generate an amplification productwherein the restriction endonuclease is one which cannot recognize or cleave the amplification product.
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Abstract
The invention provides methods and compositions for enhancing the speed and sensitivity of helicase-dependent amplification through the use of an endonuclease.
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Citations
18 Claims
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1. A method for assaying a sample, the method comprising combining a composition comprising a restriction endonuclease, a helicase, a DNA polymerase, and at least a first primer and a second primer,
with a sample containing or potentially containing a nucleic acid that hybridizes to the first and second primer, and amplifying the target sequence to generate an amplification product wherein the restriction endonuclease is one which cannot recognize or cleave the amplification product.
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9. A method for amplifying a nucleic acid, the method comprising the steps of:
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(a) treating the nucleic acid with a restriction endonuclease, (b) incubating the nucleic acid with at least a first primer and a second primer and amplifying the nucleic acid in a helicase-dependent amplification reaction to generate the amplified product, wherein the amplified product is not cleaved by the restriction endonuclease. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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Specification