Patent US2244740A successfully refreshed from Google Patents.
US6204375B1
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Methods and reagents for preserving RNA in cell and tissue samples
Assignee: Applied Biosystems LLC
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Filing: 1998-07-31
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Granted: 2001-03-20
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Expires: 2018-07-31
Abstract
Abstract This specification relates to the field of molecular biology and provides a novel method and reagent for preserving and protecting the ribonucleic acid (RNA) content of tissue samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.
Inventors
- 👤 Eric S. Lader
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-22 22:51
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6806; C12N; C12N1/00; C12N1/04; C12N15/00; C12N15/09; C12N15/10; C12N15/1003
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
80 words
A method of preserving RNA in an intact cell comprising: obtaining an intact cell; and treating the cell with an RNA preservation medium comprising at least one salt, which RNA preservation medium has a total salt concentration of between 10 g/100 ml and the saturating concentration of the salt, and which RNA preservation medium infiltrates the cell and protects RNA in the cell from nucleases; whereby said RNA is said cell is preserved at temperatures greater than −20 degree Centigrade.
Claim 2
Independent Claim
80 words
A method of preserving RNA in an intact cell comprising: obtaining an intact cell; and treating the cell with an RNA preservation medium comprising at least one salt, which RNA preservation medium has a total salt concentration of between 10 g/100 ml and the saturating concentration of the salt, and which RNA preservation medium infiltrates the cell and protects RNA in the cell from nucleases; whereby said RNA is said cell is preserved at temperatures greater than −20 degree Centigrade.
Claim 3
Dependent Claim
31 words
The method of claim 1 , wherein the RNA preservation medium precipitates the RNA in the cell along with cellular protein in the cell and renders the RNA inaccessible to nucleases.
Claim 4
Dependent Claim
13 words
The method of claim 1 , wherein the salt is a sulfate salt.
Claim 5
Dependent Claim
12 words
The method of claim 3 , wherein the salt is ammonium sulfate.
Claim 6
Dependent Claim
25 words
The method of claim 1 , wherein the salt is present in a concentration between 20 g/100 ml and the saturating concentration of the salt.
Claim 7
Dependent Claim
24 words
The method of claim 1 , wherein the salt is ammonium sulfate in a concentration of between 30 g/100 ml and 80 g/100 ml.
Claim 8
Dependent Claim
19 words
The method of claim 1 , wherein the RNA preservation medium comprises a combination of at least two salts.
Claim 9
Dependent Claim
20 words
The method of claim 7 , wherein the total salt concentration is between 20 g/100 ml and 100 g/100 ml.
Claim 10
Dependent Claim
17 words
The method of claim 1 , wherein the RNA preservation medium comprises a chelator of divalent cations.
Claim 11
Dependent Claim
14 words
The method of claim 1 , wherein the RNA preservation medium comprises a buffer.
Claim 12
Dependent Claim
19 words
The method of claim 1 , wherein the RNA preservation medium has a pH of between 4 and 8.
Claim 13
Dependent Claim
15 words
The method of claim 1 , wherein the cell is in a suspension of cells.
Claim 14
Dependent Claim
14 words
The method of claim 1 , wherein the cell is a solid tissue sample.
Claim 15
Dependent Claim
14 words
The method of claim 1 , wherein the cell is in an entire organism.
Claim 16
Dependent Claim
19 words
The method of claim 14 , wherein the organism is a pathogen within a tissue sample or other organism.
Claim 17
Dependent Claim
15 words
The method of claim 1 , further comprising the step of isolating the preserved RNA.
Claim 18
Dependent Claim
20 words
The method of claim 16 , wherein the RNA is isolated at a temperature that is greater than −20° C.
Claim 19
Dependent Claim
18 words
The method of claim 16 , wherein the cell is stored prior to the isolation of the RNA.
Claim 20
Dependent Claim
18 words
The method of claim 18 , wherein the cell is stored unfrozen at −20° C. to 45° C.
Claim 21
Dependent Claim
16 words
The method of claim 19 , wherein the cell is stored at greater than 0° C.
Claim 22
Dependent Claim
19 words
The method of claim 1 , wherein the RNA preservation medium comprises a combination of at least two salts.
Claim 23
Dependent Claim
16 words
The method of claim 1 , wherein the RNA preservation medium further comprises an organic solvent.
Claim 24
Dependent Claim
12 words
The method of claim 15 , wherein said pathogen is a virus.
Claim 25
Dependent Claim
37 words
The method of claim 1 , wherein the RNA preservation medium comprises a combination of at least two salts, which salts are present in a total salt concentration of between 20 g/100 ml and 100 g/100 ml.
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) | 2004-03-19 | 2004-09-18 | 2005-03-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-03-19 | 2008-09-18 | 2009-03-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-03-19 | 2012-09-18 | 2013-03-20 | $7700.00 | Window Closed / Lapsed |