US5126239A
Expired / Lapsed
US
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Process for detecting polymorphisms on the basis of nucleotide differences
Assignee: EIDP Inc
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Filing: 1990-03-14
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Granted: 1992-06-30
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Expires: 2010-03-14
Abstract
Abstract A process is provided for detecting polymorphisms on the basis of nucleotide differences in random segments of the nucleic acid by performing a primer extension reaction on the nucleic acids and comparing the extension reaction products. The random nucleic acid segment can be amplified by first dissociating the extension product from the template and contacting the dissociated extension product with a primer under conditions such that an amplification extension product is synthesized using the dissociated extension product as a template. Differences in the extension product are useful as markers in constructing genetic maps and as markers to distinguish or identify individuals.
Inventors
- 👤 Kenneth J. Livak
- 👤 Jan A. Rafalski
- 👤 Scott V. Tingey
- 👤 John G. Williams
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 18:19
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6844; C12Q1/6858; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
97 words
A process for detecting polymorphisms on the basis of nucleotide differences in random segments of nucleic acids comprising: (a) separately performing an extension reaction on a random segment of each of at least two nucleic acids from different sources, said reaction comprising: (i) contacting each of the nucleic acids with at least one nonspecifically targeted oligonucleotide primer of greater than 7 nucleotides under conditions such that for at least one nucleic acid a random extension product of at least one primer is synthesized; and (b) comparing the results of the separately performed random extension reactions for differences.
Claim 2
Dependent Claim
97 words
A process for detecting polymorphisms on the basis of nucleotide differences in random segments of nucleic acids comprising: (a) separately performing an extension reaction on a random segment of each of at least two nucleic acids from different sources, said reaction comprising: (i) contacting each of the nucleic acids with at least one nonspecifically targeted oligonucleotide primer of greater than 7 nucleotides under conditions such that for at least one nucleic acid a random extension product of at least one primer is synthesized; and (b) comparing the results of the separately performed random extension reactions for differences.
Claim 3
Dependent Claim
66 words
The process of claim 1, after step (i) and before step (b), further comprising the additional steps of (ii) dissociating the extension product from its complement; and (iii) amplifying the random nucleic acid segment by contacting the dissociated extension product with at least one primer of step (i) under conditions such that an amplification extension product is synthesized using the dissociated extension product as a template.
Claim 4
Dependent Claim
13 words
The process of claim 2 wherein the steps are performed at least twice.
Claim 5
Dependent Claim
22 words
The process of claim 1 or 2 using a nucleic acid polymers and nucleoside triphosphate substrates or their analogues and mixtures thereof.
Claim 6
Dependent Claim
12 words
The process of claim 1 or 2 wherein one primer is employed.
Claim 7
Dependent Claim
18 words
The process of claim 1 or 2 wherein the primer is from 9 to 10 nucleotides in length.
Claim 8
Dependent Claim
14 words
The process of claim 1 or 2 wherein at least one primer is biotinylated.
Claim 9
Dependent Claim
16 words
The process of claim 1 or 2 wherein the nucleic acids are from different individual organisms.
Claim 10
Dependent Claim
24 words
A process according to claim 4, wherein the nucleic acid polymerase is a DNA polymerase and the nucleoside triphosphate substrates are deoxyribonucleoside triphosphate substrates.
Claim 11
Dependent Claim
14 words
A process according to claim 9, wherein the DNA polymerase is a thermostable polymerase.
Claim 12
Dependent Claim
13 words
A process according to claim 10, wherein the DNA polymerase is Taq polymerase.
Claim 13
Dependent Claim
19 words
The process of claim 1 or 2, wherein the comparison is through size separation of the random extension products.
Claim 14
Dependent Claim
24 words
A process according to claim 12, wherein the size separation comparison is performed by electrophoresis through a polyacrylamide gel matrix or agarose gel matrix.
Claim 15
Dependent Claim
15 words
The process of claim 1 or 2, wherein the comparison is through nucleotide sequence determination.
Claim 16
Dependent Claim
23 words
The process of claim 1 or 2, wherein the nucleic acid source is selected from the group consisting of plants, animals and microbes.
Claim 17
Dependent Claim
14 words
The process of claim 1 or 2 wherein the nucleic acid source is human.
Claim 18
Dependent Claim
25 words
The process of claim 1 or 2 wherein a difference in the random extension products is used as a marker to construct a genetic map.
Claim 19
Dependent Claim
25 words
The process of claim 1 or 2 wherein a difference in the random extension products is used as a marker to distinguish or identify individuals.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-06-30 | 1995-12-30 | 1996-06-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-06-30 | 1999-12-30 | 2000-06-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-06-30 | 2003-12-30 | 2004-06-30 | $7700.00 | Window Closed / Lapsed |