US4921805A
Expired / Lapsed
US
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Nucleic acid capture method
Assignee: Life Technologies Inc
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Filing: 1989-09-29
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Granted: 1990-05-01
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Expires: 2009-09-29
Abstract
Abstract A nucleic acid capture reagent which comprises one or more molecules that are capable of intercalation into nucleic acids and are attached to a solid support via a molecular linker. This capture reagent is useful for the separation and isolation of nucleic acids from complex unpurified biological solutions such as serum, sputum, blood, and urine. The resulting capture reagent-nucleic acid complexes are easily isolated from the sample solution by mechanical or magnetic means and the bound nucleic acids are released by simple chemical treatment.
Inventors
- 👤 Gulilat Gebeyehu
- 👤 Leonard Klevan
- 👤 John D. Harding
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 08:46
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6806; C12Q1/6813; C12Q1/6834; Y; Y10; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375; Y10T436/255
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
127 words
A method of separating nucleic acids from an unpurified biological sample comprising: (a) contacting an unpurified biological sample with a capture reagent, wherein said capture reagent comprises: (i) a solid support; (ii) a linker molecule comprising spermine or spermidine bound to said solid support; and (iii) a methidium moiety, bound to said linker; wherein said methidium moiety is able to bind to the nucleic acids in said biological sample independent of the salt concentration of said sample; and wherein a sample-capture reagent mixture is formed thereby; (b) insulating said sample-capture reagent mixture for a time and under conditions sufficient to allow said capture reagent to bind to nucleic acids in said sample to form a capture reagent-nucleic acid complex; and (c) isolating said complex from said mixture.
Claim 2
Independent Claim
127 words
A method of separating nucleic acids from an unpurified biological sample comprising: (a) contacting an unpurified biological sample with a capture reagent, wherein said capture reagent comprises: (i) a solid support; (ii) a linker molecule comprising spermine or spermidine bound to said solid support; and (iii) a methidium moiety, bound to said linker; wherein said methidium moiety is able to bind to the nucleic acids in said biological sample independent of the salt concentration of said sample; and wherein a sample-capture reagent mixture is formed thereby; (b) insulating said sample-capture reagent mixture for a time and under conditions sufficient to allow said capture reagent to bind to nucleic acids in said sample to form a capture reagent-nucleic acid complex; and (c) isolating said complex from said mixture.
Claim 3
Dependent Claim
24 words
The method of claim 1 wherein said isolating is performed by the method selected from the group consisting of centrifugation, filtration, and magnetic separation.
Claim 4
Dependent Claim
31 words
The method of claim 1 further comprising: (a) treating said isolated complex to effect release of said nucleic acid therefrom; and (b) separating said released nucleic acids from said isolated complex.
Claim 5
Dependent Claim
18 words
The method of claim 3 wherein step (a) comprises treating said capture reagent-nucleic acid complex with dilute alkali.
Claim 6
Dependent Claim
13 words
The method of claim 1 wherein said solid support is a beaded material.
Claim 7
Dependent Claim
21 words
The method of claim 5 wherein said beaded material is selected from the group consisting of sepharose, agarose, or magnetic beads.
Claim 8
Dependent Claim
12 words
The method of claim 1 wherein said solid support is a polymer.
Claim 9
Dependent Claim
28 words
The method of claim 7 wherein the form of said polymeric solid support is selected from the group consisting of a tube, a dipstick, or a microtiter plate.
Claim 10
Dependent Claim
12 words
The method of claim 1 wherein said solid support is a membrane.
Claim 11
Dependent Claim
13 words
The method of claim reagent of claim 9 wherein said membrane comprises nylon.
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) | 1993-04-30 | 1993-10-30 | 1994-05-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-04-30 | 1997-10-30 | 1998-05-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-04-30 | 2001-10-30 | 2002-05-01 | $7700.00 | Window Closed / Lapsed |