US4767700A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Detection of particular nucleotide sequences
Assignee: Beckman Research Institute of City of Hope
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Filing: 1985-02-15
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Granted: 1988-08-30
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Expires: 2005-02-15
Abstract
Abstract A process for the detection of a particular polynucleotide sequence in a polynucleotide-containing test sample comprising forming an agarose gel having a depression therein, adding the test sample to said depression, putting agarose solution in the depression and permitting it to gel thereby to encase the test sample in agarose gel, denaturing the polynucleotide in the test sample within the agarose gel, under hybridization conditions contacting the test sample-containing gel with an oligonucleotide or polynucleotide probe having a sequence complementary with a sequence for which the sample is being tested, and determining the extent of hybridization.
Inventors
- 👤 R. Bruce Wallace
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 23:06
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/5436; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6827; C12Q1/6876; C12Q1/6881; C12Q1/6883; Y; Y10; Y10S; Y10S435/00; Y10S435/803; Y10S436/00; Y10S436/811
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
132 words
A process for the detection of a particular polynucleotide sequence in a polynucleotide-containing test sample consisting essentially of forming an agarose gel having a depression therein, adding the test sample to said depression, putting agarose solution in the depression and permitting it to gel thereby encasing the test sample in agarose gel, denturing the polynucleotide in the test sample within the agarose gel, contacting the test sample-containing gel with an oligonucleotide or polynucleotide probe having a sequence complementary with a sequence for which the sample is being tested under hybridization conditions, said probe having at least 23 nucleotides, said probe carrying a readable label, and determining the extent of hybridization, the extent of hybridization being determined by reading the label and, wherein after hybridization the gel is washed to remove non-hybridized probe.
Claim 2
Dependent Claim
132 words
A process for the detection of a particular polynucleotide sequence in a polynucleotide-containing test sample consisting essentially of forming an agarose gel having a depression therein, adding the test sample to said depression, putting agarose solution in the depression and permitting it to gel thereby encasing the test sample in agarose gel, denturing the polynucleotide in the test sample within the agarose gel, contacting the test sample-containing gel with an oligonucleotide or polynucleotide probe having a sequence complementary with a sequence for which the sample is being tested under hybridization conditions, said probe having at least 23 nucleotides, said probe carrying a readable label, and determining the extent of hybridization, the extent of hybridization being determined by reading the label and, wherein after hybridization the gel is washed to remove non-hybridized probe.
Claim 3
Dependent Claim
51 words
A proces according to claim 1, wherein the hybridization is conducted in 5×SSPE, wherein SSPE comprises 0.18M NaCl, 10 mM, (Na 1 .5) PO 4 , 1 mM Na 2 EDTA, pH of 7.0, 0.1% SDS and 10 μg/m; of sonicated, denatured E. coli DNA for 3 hours at 60° C.
Claim 4
Dependent Claim
13 words
A process according to claim 1 wherein the label is a radioactive isotope.
Claim 5
Dependent Claim
11 words
A process according to claim 1 wherein the label is biotin.
Claim 6
Dependent Claim
16 words
A process according to claim 1, wherein the encased test sample is dried prior to denaturing.
Claim 7
Dependent Claim
12 words
A process according to claim 1, wherein the probe is an oligonucleotide.
Claim 8
Dependent Claim
30 words
A process according to claim 1, wherein the probe is a polynucleotide sequence which distinguishes between different sequences in test samples from individuals suspected of having a particular genetic condition.
Claim 9
Dependent Claim
14 words
A process according to claim 7, wherein the genetic condition is sickle cell anemia.
Claim 10
Dependent Claim
12 words
A process according to claim 7, wherein the genetic condition is thalassemia.
References Cited (Backward Citations)
US4395486A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-08-30 | 1992-02-29 | 1992-08-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-08-30 | 1996-02-29 | 1996-08-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-08-30 | 2000-02-29 | 2000-08-30 | $7700.00 | Window Closed / Lapsed |