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US 9,822,401 B2Granted
Methods and systems for nucleic acid amplification
AssigneeGenapsys, Inc.
InventorFlorian Oberstrass
Life of this patent
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Abstract
The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.
Claims41
A method for amplifying a nucleic acid sample, comprising:
- a) providing a support comprising a first primer and a second primer;
- b) coupling the first primer to a first single-stranded nucleic acid molecule that is derived from the nucleic acid sample;
- c) performing a first primer extension reaction using the first primer to generate a first double-stranded nucleic acid molecule comprising the first single-stranded nucleic acid molecule and a second single-stranded nucleic acid molecule that is complementary to at least a portion of the first single-stranded nucleic acid molecule;
- d) at least partially denaturing the first double-stranded nucleic acid molecule by binding an invader species to at least a portion of the first or second single-stranded nucleic acid molecule, wherein the invader species is different from the second primer, which binding exposes a segment of the first single-stranded nucleic acid molecule that is complementary to the second primer;
- and
- e) coupling the second primer to the segment of the first single-stranded nucleic acid molecule and performing a second primer extension reaction using the second 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 second primer extension reaction separates the second single-stranded nucleic acid molecule from the first single-stranded nucleic acid molecule.
3 more elements
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 double-stranded nucleic acid molecule is derived from the nucleic acid sample, and wherein the second-single-stranded nucleic acid molecule comprises a nucleic acid sequence at least partially complementary to said first primer;
- 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 or second single-stranded nucleic acid molecule, which binding exposes a segment of the first single-stranded nucleic acid molecule that is complementary to a second primer separate from said first primer;
- d) coupling the second primer to the segment of the first single-stranded nucleic acid molecule and performing a first primer extension reaction using the second 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;
- and
- e) coupling the first primer to the separated second-single stranded molecule and performing a second primer extension reaction using the first primer to generate a third double-stranded nucleic acid molecule comprising the second single-stranded nucleic acid molecule and a fourth single-stranded nucleic acid molecule.
3 more elements
A method for amplifying a nucleic acid sample, comprising:
- a) providing a support comprising a first primer;
- b) coupling the first primer to a first single-stranded nucleic acid molecule that is derived from the nucleic acid sample;
- c) performing a first primer extension reaction using the first primer to generate a first double-stranded nucleic acid molecule comprising the first single-stranded nucleic acid molecule and a second single-stranded nucleic acid molecule that is complementary to at least a portion of the first single-stranded nucleic acid molecule;
- d) at least partially denaturing the first double-stranded nucleic acid molecule by binding an invader species free from said support to at least a portion of the first or second single-stranded nucleic acid molecule, which binding exposes a segment of the first or second single-stranded nucleic acid molecule that is complementary to a second primer;
- and
- e) coupling the second primer to the segment of the first or second single-stranded nucleic acid molecule and performing a second primer extension reaction using the second primer to generate a second double-stranded nucleic acid molecule comprising the first or second single-stranded nucleic acid molecule and a third single-stranded nucleic acid molecule that is complementary to at least a portion of the first or second single-stranded nucleic acid molecule, wherein the second primer extension reaction separates the first single-stranded nucleic acid molecule from the second single-stranded nucleic acid molecule.
3 more elements
Dependencies are inferred from the claim text.