US4921952A
Expired / Lapsed
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Nucleic acid isolation process
Assignee: United States Department of Energy
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Filing: 1989-04-20
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Granted: 1990-05-01
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Expires: 2009-04-20
Abstract
Abstract A method is provided for isolating DNA from eukaryotic cell and flow sorted chromosomes. When DNA is removed from chromosome and cell structure, detergent and proteolytic digestion products remain with the DNA. These products can be removed with organic extraction, but the process steps associated with organic extraction reduce the size of DNA fragments available for experimental use. The present process removes the waste products by dialyzing a solution containing the DNA against a solution containing polyethylene glycol (PEG). The waste products dialyze into the PEG leaving isolated DNA. The remaining DNA has been prepared with fragments containing more than 160 kb. The isolated DNA has been used in conventional protocols without affect on the protocol.
Inventors
- 👤 Jonathan L. Longmire
- 👤 Annette K. Lewis
- 👤 Carl E. Hildebrand
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 10:08
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1003; C12Q; C12Q1/00; C12Q1/68; C12Q1/6806; Y; Y10; Y10S; Y10S435/00; Y10S435/803
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
84 words
In a process for preparing DNA from cellular materials for use in genetic studies of eukaryotic systems, a process for isolating DNA fragments from proteolytic digestion products and detergent products in a solution with said DNA fragments produced in stripping undesired cellular constituents from said DNA, wherein the improvement comprises the step of dialyzing a solution containing said DNA fragments, detergent products, and proteolytic digestion products against a solution containing PEG for a time effective to yield DNA sufficiently pure for said genetic studies.
Claim 2
Dependent Claim
84 words
In a process for preparing DNA from cellular materials for use in genetic studies of eukaryotic systems, a process for isolating DNA fragments from proteolytic digestion products and detergent products in a solution with said DNA fragments produced in stripping undesired cellular constituents from said DNA, wherein the improvement comprises the step of dialyzing a solution containing said DNA fragments, detergent products, and proteolytic digestion products against a solution containing PEG for a time effective to yield DNA sufficiently pure for said genetic studies.
Claim 3
Dependent Claim
16 words
A process according to claim 1, wherein said solution containing PEG is about 20% (w/v) PEG.
Claim 4
Dependent Claim
21 words
A process according to claim 1, further including the step of performing periodic changes of said PEG solution during said dialysis.
Claim 5
Dependent Claim
67 words
In a process for preparing DNA from cellular materials for use in genetic studies of eukaryotic systems, a process for isolating DNA fragments from proteolytic digestion products and detergent products in a solution with said DNA fragments produced in stripping undesired cellular constituents from said DNA, consisting essentially of dialyzing a solution containing said DNA fragments, detergent products, and proteolytic digestion products against a solution containing PEG.
Claim 6
Dependent Claim
67 words
In a process for preparing DNA from cellular materials for use in genetic studies of eukaryotic systems, a process for isolating DNA fragments from proteolytic digestion products and detergent products in a solution with said DNA fragments produced in stripping undesired cellular constituents from said DNA, consisting essentially of dialyzing a solution containing said DNA fragments, detergent products, and proteolytic digestion products against a solution containing PEG.
Claim 7
Dependent Claim
16 words
A process according to claim 4, wherein said solution containing PEG is about 20% (w/v) PEG.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 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 |