Patent DE2748858A1 successfully refreshed from Google Patents.
US6248535B1
Expired / Lapsed
US
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Method for isolation of RNA from formalin-fixed paraffin-embedded tissue specimens
Assignee: University of Southern California
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Filing: 1999-12-20
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Granted: 2001-06-19
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Expires: 2019-12-20
Abstract
Abstract Methods are disclosed for rapid, reliable and simple isolation of RNA from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used to monitor gene expression levels. The tissue sample can be a tumor or other pathological tissue.
Inventors
- 👤 Kathleen Danenberg
- 👤 Peter V. Danenberg
- 👤 Steven Swenson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 09:39
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1003; C12Q; C12Q2600/00; C12Q2600/158
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
69 words
A method for recovering DNA, RNA or proteins from a formalin-fixed paraffin-embedded biological tissue sample comprising: deparaffinizing the sample; heating the sample in a chaotropic solution comprising an effective concentration of a guanidinium compound to a temperature in the range of about 60 to about 100° C. for a time period of about 30 to about 120 minutes; and recovering said RNA, DNA or protein from said chaotropic solution.
Claim 2
Independent Claim
69 words
A method for recovering DNA, RNA or proteins from a formalin-fixed paraffin-embedded biological tissue sample comprising: deparaffinizing the sample; heating the sample in a chaotropic solution comprising an effective concentration of a guanidinium compound to a temperature in the range of about 60 to about 100° C. for a time period of about 30 to about 120 minutes; and recovering said RNA, DNA or protein from said chaotropic solution.
Claim 3
Dependent Claim
13 words
The method of claim 1 further comprising rehydrating the deparaffinized sample before heating.
Claim 4
Dependent Claim
12 words
The method of claim 2 further comprising homogenizing said sample before heating.
Claim 5
Dependent Claim
30 words
The method of claim 3 for recovering RNA from a biological tissue sample wherein said RNA is recovered by extraction from said chaotropic solution with a water insoluble organic solvent.
Claim 6
Dependent Claim
13 words
The method of claim 4 wherein said water insoluble organic solvent comprises chloroform.
Claim 7
Dependent Claim
10 words
The method of claim 5 further comprising purifying said RNA.
Claim 8
Dependent Claim
13 words
The method of claim 6 wherein said RNA is purified by ethanol precipitation.
Claim 9
Dependent Claim
20 words
The method of claim 1 for recovering RNA wherein said time period is from about 10 to about 60 minutes.
Claim 10
Dependent Claim
17 words
The method of claim 8 wherein said time period is from about 30 to about 60 minutes.
Claim 11
Dependent Claim
15 words
The method of claim 1 for recovering RNA wherein said guanidinium compound is guanidinium hydrochloride.
Claim 12
Dependent Claim
15 words
The method of claim 1 for recovering RNA wherein said guanidinium compound is guanidinium isothiocyanate.
Claim 13
Dependent Claim
27 words
The method of claim 11 wherein the concentration of said guanidinium isothiocyanate in the chaotropic solution is present in a concentration of about 2 to about 5M.
Claim 14
Dependent Claim
15 words
The method of claim 11 wherein said chaotropic solution has a pH of about 3-6.
Claim 15
Dependent Claim
17 words
The method of claim 1 for recovering RNA wherein said chaotropic solution further comprises a reducing agent.
Claim 16
Dependent Claim
11 words
The method of claim 14 wherein said reducing agent is β-mercaptoethanol.
Claim 17
Dependent Claim
11 words
The method of claim 14 wherein said reducing agent is dithiothreitol.
Claim 18
Dependent Claim
19 words
The method of claim 1 wherein said RNA is used to determine the level of expression of a gene.
Claim 19
Independent Claim
63 words
A method for isolating RNA from a formalin-fixed paraffin-embedded biological tissue sample comprising: deparaffinizing the sample; heating the sample in a chaotropic solution comprising an effective concentration of a guanidinium compound to a temperature in the range of about 75 to about 100° C. for a time period of about 30 to about 60 minutes; and recovering said RNA from said chaotropic solution.
Claim 20
Independent Claim
63 words
A method for isolating RNA from a formalin-fixed paraffin-embedded biological tissue sample comprising: deparaffinizing the sample; heating the sample in a chaotropic solution comprising an effective concentration of a guanidinium compound to a temperature in the range of about 75 to about 100° C. for a time period of about 30 to about 60 minutes; and recovering said RNA from said chaotropic solution.
Claim 21
Dependent Claim
13 words
The method of claim 18 further comprising rehydrating the deparaffinized sample before heating.
Claim 22
Dependent Claim
12 words
The method of claim 19 further comprising homogenizing said sample before heating.
Claim 23
Dependent Claim
30 words
The method of claim 20 for recovering RNA from a biological tissue sample wherein said RNA is recovered by extraction from said chaotropic solution with a water insoluble organic solvent.
Claim 24
Dependent Claim
13 words
The method of claim 21 wherein said water insoluble organic solvent comprises chloroform.
Claim 25
Dependent Claim
10 words
The method of claim 22 further comprising purifying said RNA.
Claim 26
Dependent Claim
13 words
The method of claim 23 wherein said RNA is purified by ethanol precipitation.
Claim 27
Dependent Claim
15 words
The method of claim 18 for recovering RNA wherein said guanidinium compound is guanidinium hydrochloride.
Claim 28
Dependent Claim
15 words
The method of claim 18 for recovering RNA wherein said guanidinium compound is guanidinium isothiocyanate.
Claim 29
Dependent Claim
22 words
The method of claim 26 wherein the concentration of said guanidinium isothiocyanate in the chaotropic solution is about 2 to about 5M.
Claim 30
Dependent Claim
15 words
The method of claim 26 wherein said chaotropic solution has a pH of about 3-6.
Claim 31
Dependent Claim
17 words
The method of claim 18 for recovering RNA wherein said chaotropic solution further comprises a reducing agent.
Claim 32
Dependent Claim
11 words
The method of claim 29 wherein said reducing agent is β-mercaptoethanol.
Claim 33
Dependent Claim
11 words
The method of claim 29 wherein said reducing agent is dithiothreitol.
Claim 34
Dependent Claim
19 words
The method of claim 18 wherein said RNA is used to determine the level of expression of a gene.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2004-06-18 | 2004-12-18 | 2005-06-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-06-18 | 2008-12-18 | 2009-06-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-06-18 | 2012-12-18 | 2013-06-19 | $7700.00 | Window Closed / Lapsed |