Methods and systems for nucleic acid amplification
First Claim
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1. A method for amplifying a nucleic acid sample, comprising:
- a) providing a support comprising a first primer;
b) providing a first double-stranded nucleic acid molecule comprising a first single-stranded nucleic acid molecule and a second single-stranded nucleic acid molecule that is at least partially complementary to said first single-stranded nucleic acid molecule, wherein the first double-stranded nucleic acid molecule is derived from the nucleic acid sample;
c) at least partially denaturing the first double-stranded nucleic acid molecule by binding a first invader species free from said support to at least a portion of the first single-stranded nucleic acid molecule or at least a portion of the second single-stranded nucleic acid molecule, which binding exposes a first segment of the first single-stranded nucleic acid molecule that is complementary to the first primer; and
d) coupling the first primer to the first segment of the first single-stranded nucleic acid molecule and performing a first primer extension reaction using the first primer to generate a second double-stranded nucleic acid molecule comprising the first single-stranded nucleic acid molecule and a third single-stranded nucleic acid molecule that is complementary to at least a portion of the first single-stranded nucleic acid molecule, wherein the first primer extension reaction separates the second single-stranded nucleic acid molecule from the first single-stranded nucleic acid molecule.
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Abstract
The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.
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Citations
20 Claims
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1. A method for amplifying a nucleic acid sample, comprising:
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a) providing a support comprising a first primer; b) providing a first double-stranded nucleic acid molecule comprising a first single-stranded nucleic acid molecule and a second single-stranded nucleic acid molecule that is at least partially complementary to said first single-stranded nucleic acid molecule, wherein the first double-stranded nucleic acid molecule is derived from the nucleic acid sample; c) at least partially denaturing the first double-stranded nucleic acid molecule by binding a first invader species free from said support to at least a portion of the first single-stranded nucleic acid molecule or at least a portion of the second single-stranded nucleic acid molecule, which binding exposes a first segment of the first single-stranded nucleic acid molecule that is complementary to the first primer; and d) coupling the first primer to the first segment of the first single-stranded nucleic acid molecule and performing a first primer extension reaction using the first primer to generate a second double-stranded nucleic acid molecule comprising the first single-stranded nucleic acid molecule and a third single-stranded nucleic acid molecule that is complementary to at least a portion of the first single-stranded nucleic acid molecule, wherein the first primer extension reaction separates the second single-stranded nucleic acid molecule from the first single-stranded nucleic acid molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification