US4833239A
Expired / Lapsed
US
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Method for the isolation and purification of DNA molecules
Assignee: Genetics Institute LLC
•
Filing: 1987-01-29
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Granted: 1989-05-23
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Expires: 2007-01-29
Abstract
Abstract A method for isolating and purifying nucleic acids from eukaryotic and prokaryotic cell cultures or viral cell cultures.
Inventors
- 👤 David A. DeBonville
- 👤 Gerard E. Riedel
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 15:52
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1003
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
153 words
A method for isolating and purifying DNA molecules from cell culture medium of the type in which: (a) cells in the culture are concentrated apart from major contaminants in the media; (b) the cells are lysed and deproteinated; (c) cellular debris is eliminated; (d) the DNA is extracted by precipitation; (e) the DNA is washed to remove contaminants; and (f) the precipitated DNA is isolated by elution; wherein the improvement comprises performing said lysing and deproteinating in one step with a stable single phase aqueous reagent composition comprising: (i) about 1.6 to 3.2M potassium acetate; (ii) about 5 to 15% by weight phenol; (iii) about 5 to 15% by weight chloroform; and (iv) glacial acetic acid in an amount such that the weight ratio of potassium acetate to acetic acid is 3:2, wherein the weight ratio of (i) to the combination of (ii) and (iii) is in the range of 4:1 to 4:3.
Claim 2
Independent Claim
153 words
A method for isolating and purifying DNA molecules from cell culture medium of the type in which: (a) cells in the culture are concentrated apart from major contaminants in the media; (b) the cells are lysed and deproteinated; (c) cellular debris is eliminated; (d) the DNA is extracted by precipitation; (e) the DNA is washed to remove contaminants; and (f) the precipitated DNA is isolated by elution; wherein the improvement comprises performing said lysing and deproteinating in one step with a stable single phase aqueous reagent composition comprising: (i) about 1.6 to 3.2M potassium acetate; (ii) about 5 to 15% by weight phenol; (iii) about 5 to 15% by weight chloroform; and (iv) glacial acetic acid in an amount such that the weight ratio of potassium acetate to acetic acid is 3:2, wherein the weight ratio of (i) to the combination of (ii) and (iii) is in the range of 4:1 to 4:3.
Claim 3
Dependent Claim
16 words
The method according to claim 1 wherein DNA comprises double-stranded plasmid DNA or single-stranded bacteriophage DNA.
Claim 4
Dependent Claim
11 words
The method according to claim 2 wherein said bacteriophage is M13.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-05-22 | 1992-11-21 | 1993-05-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-05-22 | 1996-11-21 | 1997-05-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-05-22 | 2000-11-21 | 2001-05-23 | $7700.00 | Window Closed / Lapsed |