US4483920A
Expired / Lapsed
US
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Immobilization of message RNA directly from cells onto filter material
Assignee: Hahnemann University
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Filing: 1982-05-17
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Granted: 1984-11-20
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Expires: 2002-05-17
Abstract
Abstract Message RNA is immobilized directly from cells onto filter material. Immobilization is carried out by solubilizing cellular components with a chaotropic salt, passing the resultant solubilized cellular components through a filter which selectively binds message RNA and baking the filter containing bound message RNA. The chaotropic salt is preferably sodium iodide, potassium iodide or sodium perchlorate. Prior to solubilizing, the cells may be washed and lysed. The bound message RNA can be hybridized to a labeled probe and the amount of message RNA measured. Prior to baking, the filter containing bound RNA may be incubated in a solution which acelylates basic protein and other molecules which might interfere with molecular hybridization.
Inventors
- 👤 David Gillespie
- 👤 Isadore Brodsky
- 👤 Joel Bresser
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 06:32
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6806
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
84 words
A method of immobilizing message RNA from cells, comprising the steps of: A. Solubilizing cells containing message RNA by mixing the cells with an aqueous solution of a chaotropic salt selected from the group consisting of sodium iodide, potassium iodide, and sodium perchlorate; B. thereafter filtering the mixture resulting from step (A) through a filter which selectively binds and immobilizes said message RNA; and C. baking by heating the filter resulting from step (B) to further bond said message RNA to the filter material.
Claim 2
Independent Claim
84 words
A method of immobilizing message RNA from cells, comprising the steps of: A. Solubilizing cells containing message RNA by mixing the cells with an aqueous solution of a chaotropic salt selected from the group consisting of sodium iodide, potassium iodide, and sodium perchlorate; B. thereafter filtering the mixture resulting from step (A) through a filter which selectively binds and immobilizes said message RNA; and C. baking by heating the filter resulting from step (B) to further bond said message RNA to the filter material.
Claim 3
Dependent Claim
11 words
The method of claim 1, wherein said filter material is nitrocellulose.
Claim 4
Dependent Claim
116 words
A process for measuring quantities of a specific message RNA from cells, comprising the steps of: A. Solubilizing cells containing message RNA by mixing the cells with an aqueous solution of a chaotropic salt selected from the group consisting of sodium iodide, potassium iodide, and sodium perchlorate; B. thereafter filtering the mixture resulting from step (A) through a filter which selectively binds and immobilizes said message RNA; C. baking by heating filter resulting from step (B) to further bond said message RNA to the filter material; and D. hybridizing said bonded message RNA resulting from step (C) to a labeled probe and measuring the amount of said message RNA based on the extent of said hybridization.
Claim 5
Dependent Claim
116 words
A process for measuring quantities of a specific message RNA from cells, comprising the steps of: A. Solubilizing cells containing message RNA by mixing the cells with an aqueous solution of a chaotropic salt selected from the group consisting of sodium iodide, potassium iodide, and sodium perchlorate; B. thereafter filtering the mixture resulting from step (A) through a filter which selectively binds and immobilizes said message RNA; C. baking by heating filter resulting from step (B) to further bond said message RNA to the filter material; and D. hybridizing said bonded message RNA resulting from step (C) to a labeled probe and measuring the amount of said message RNA based on the extent of said hybridization.
Claim 6
Dependent Claim
11 words
The method of claim 3, wherein said filter material is nitrocellulose.
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) | 1987-11-20 | 1988-05-21 | 1988-11-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-11-20 | 1992-05-21 | 1992-11-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-11-20 | 1996-05-21 | 1996-11-20 | $7700.00 | Window Closed / Lapsed |