US5498523A
Expired / Lapsed
US
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DNA sequencing with pyrophosphatase
Assignee: Harvard University
•
Filing: 1994-07-14
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Granted: 1996-03-12
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Expires: 2014-07-14
Abstract
Abstract A kit or solution for use in extension of an oligonucleotide primer having a first single-stranded region on a template molecule having a second single-stranded region homologous to the first single-stranded region, comprising a first agent able to cause extension of the first single-stranded region of the primer on the second single-stranded region of the template in a reaction mixture, and a second agent able to reduce the amount of pyrophosphate in the reaction mixture below the amount produced during the extension in the absence of the second agent.
Inventors
- 👤 Stanley Tabor
- 👤 Charles C. Richardson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 20:53
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/16; C12Q; C12Q1/00; C12Q1/68; C12Q1/6844; C12Q1/686; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
20 words
A solution for use in a DNA sequencing reaction, comprising a DNA polymerase, a chain terminating agent, and a pyrophosphatase.
Claim 2
Dependent Claim
20 words
A solution for use in a DNA sequencing reaction, comprising a DNA polymerase, a chain terminating agent, and a pyrophosphatase.
Claim 3
Dependent Claim
19 words
The solution of claim 1, wherein said pyrophosphatase retains activity at a temperature between 60° C. and 95° C.
Claim 4
Dependent Claim
31 words
An improved method for performing DNA sequencing reactions, including providing a DNA polymerase, the improvement comprising: performing the DNA sequencing reaction in the presence of an added pyrophosphatase and DNA polymerase.
Claim 5
Dependent Claim
31 words
An improved method for performing DNA sequencing reactions, including providing a DNA polymerase, the improvement comprising: performing the DNA sequencing reaction in the presence of an added pyrophosphatase and DNA polymerase.
Claim 6
Dependent Claim
17 words
The method of claim 3, wherein said method further comprises the step of providing a dideoxynucleoside triphosphate.
Claim 7
Dependent Claim
21 words
The solution of claim 1 wherein said DNA polymerase is chosen from T7 DNA polymerase, reverse transcriptase, and Φ29 DNA polymerase.
Claim 8
Dependent Claim
21 words
The method of claim 3 wherein said DNA polymerase is chosen from T7 DNA polymerase, reverse transcriptase, and Φ29 DNA polymerase.
Claim 9
Dependent Claim
14 words
The solution of claim 1, wherein said chain terminating agent is a dideoxynucleoside triphosphate.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-03-12 | 1999-09-11 | 2000-03-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-03-12 | 2003-09-11 | 2004-03-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-03-12 | 2007-09-11 | 2008-03-12 | $7700.00 | Window Closed / Lapsed |