US5091310A
Expired / Lapsed
US
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Structure-independent dna amplification by the polymerase chain reaction
Assignee: Roche Molecular Systems Inc
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Filing: 1988-09-23
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Granted: 1992-02-25
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Expires: 2008-09-23
Abstract
Abstract Structure-independent amplification of DNA by the polymerase chain reaction can be achieved by incorporation of 7-deaza-2'-deoxyguanosine-5'-triphosphate into the amplified DNA.
Inventors
- 👤 Michael A. Innis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 09:01
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6813; C12Q1/6816; C12Q1/6844; C12Q1/686; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S436/00; Y10S436/807; Y10S436/808
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
120 words
A method for structure-independent amplification of DNA by the polymerase chain reaction, said method comprising: (a) treating the DNA under hybridizing conditions with a pair of oligonucleotide primers, a DNA polymerase, dATP, dCTP, TTP, and c 7 dGTP such that an extension product of each oligonucleotide primer is formed that is complementary to the DNA, wherein the extension product of a first primer of said primer pair, when separated from its template, can serve as a template for synthesis of the extension product of a second primer of said pair; (b) separating the extension products from the templates on which the extension products were synthesized; and (c) repeating steps (a) and (b) on the extension products produced in step (b).
Claim 2
Independent Claim
120 words
A method for structure-independent amplification of DNA by the polymerase chain reaction, said method comprising: (a) treating the DNA under hybridizing conditions with a pair of oligonucleotide primers, a DNA polymerase, dATP, dCTP, TTP, and c 7 dGTP such that an extension product of each oligonucleotide primer is formed that is complementary to the DNA, wherein the extension product of a first primer of said primer pair, when separated from its template, can serve as a template for synthesis of the extension product of a second primer of said pair; (b) separating the extension products from the templates on which the extension products were synthesized; and (c) repeating steps (a) and (b) on the extension products produced in step (b).
Claim 3
Dependent Claim
13 words
The method of claim 1, wherein dGTP is also present in step (a).
Claim 4
Dependent Claim
14 words
The method of claim 1, wherein said DNA polymerase is Thermus aquaticus DNA polymerase.
Claim 5
Dependent Claim
30 words
The method of claim 1, wherein said DNA was produced from RNA by: (a) synthesizing a cDNA from said RNA; and (b) synthesizing a double stranded DNA from said cDNA.
Claim 6
Dependent Claim
24 words
The method of claim 1, wherein the concentration of dATP, TTP, dCTP, and c 7 dGTP is each between 10 μM and 1.5 mM.
Claim 7
Dependent Claim
23 words
The method of claim 2, wherein said dGTP and said c 7 dGTP are present at a ratio of 1:3 dGTP:c 7 dGTP.
Claim 8
Dependent Claim
14 words
The method of claim 2, wherein said DNA polymerase is Thermus aquaticus DNA polymerase.
Claim 9
Dependent Claim
14 words
The method of claim 4, wherein said DNA polymerase is Thermus aquaticus DNA polymerase.
Claim 10
Dependent Claim
18 words
The method of claim 4, wherein c 7 dGTP is incorporated in said cDNA and double stranded DNA.
Claim 11
Dependent Claim
14 words
The method of claim 5, wherein said DNA polymerase is Thermus aquaticus DNA polymerase.
Claim 12
Dependent Claim
13 words
The method of claim 10, wherein dGTP is also present in step (a).
Claim 13
Dependent Claim
23 words
The method of claim 11, wherein said dGTP and said c 7 dGTP are present at a ratio of 1:3 dGTP:c 7 dGTP.
Claim 14
Dependent Claim
17 words
The method of claim 12, wherein said dGTP is present at a concentration of about 50 μM.
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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) | 1995-02-24 | 1995-08-26 | 1996-02-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-02-24 | 1999-08-26 | 2000-02-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-02-24 | 2003-08-26 | 2004-02-25 | $7700.00 | Window Closed / Lapsed |