US5234824A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Rapid purification of DNA
Assignee: Specialty Laboratories Inc
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Filing: 1992-06-02
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Granted: 1993-08-10
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Expires: 2012-06-02
Abstract
Abstract The present invention provides a method for rapidly obtaining substantially pure DNA from a biological sample containing cells. The method involves gently lysing the membranes of the cells to yield a lysate containing genomic DNA in a high molecular weight form. The lysate is moved through a porous filter to selectively trap the high molecular weight DNA on the filter. The DNA is released from the filter using an aqueous solution to form a solution containing substantially purified DNA, from which the DNA may analyzed or recovered.
Inventors
- 👤 Kary B. Mullis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 17:55
CPC Classifications
C; C12; C12N; C12N1/00; C12N1/06; C12N15/00; C12N15/09; C12N15/10; C12N15/1003; C12N15/1017; Y; Y10; Y10S; Y10S435/00; Y10S435/803; Y10S435/82
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
129 words
A method for rapidly obtaining substantially pure DNA from blood cells comprising the steps of: (a) avoiding high shear forces by gently lysing the membranes of said cells for about from two to about fifty minutes with a solution comprising a detergent and a viscosity-increasing agent to release the contents of the cells as a lysate containing DNA, a portion of said DNA having a molecular weight sufficiently large to be selectively trapped on a membrane filter; and (b) filtering said lysate through a membrane filter to selectively trap said portion of DNA on the filter, wherein said filter comprises a porous substrate having a surface that reversibly and specifically traps said portion of DNA, said filter having a pore size from about 0.2 microns to about 0.8 microns.
Claim 2
Independent Claim
129 words
A method for rapidly obtaining substantially pure DNA from blood cells comprising the steps of: (a) avoiding high shear forces by gently lysing the membranes of said cells for about from two to about fifty minutes with a solution comprising a detergent and a viscosity-increasing agent to release the contents of the cells as a lysate containing DNA, a portion of said DNA having a molecular weight sufficiently large to be selectively trapped on a membrane filter; and (b) filtering said lysate through a membrane filter to selectively trap said portion of DNA on the filter, wherein said filter comprises a porous substrate having a surface that reversibly and specifically traps said portion of DNA, said filter having a pore size from about 0.2 microns to about 0.8 microns.
Claim 3
Dependent Claim
86 words
The method of claim 1 further comprising the steps of: (a) releasing said portion of DNA from said filter wherein said step of releasing comprises immersing said membrane filter for a period of from about 5 minutes to about 15 minutes in a solution of substantially pure water at a temperature of about 100° C. to release said portion of DNA trapped on said membrane filter into said solution to form a substantially purified solution of DNA; and (b) analyzing said substantially purified solution of DNA.
Claim 4
Dependent Claim
23 words
The method of claim 2 further comprising recovering said portion of DNA in a substantially purified form from said solution of distilled water.
Claim 5
Dependent Claim
35 words
The method of claim 2 wherein said solution of substantially purified water further comprises divalent cations selected from the group consisting of magnesium and calcium, wherein the cation concentration is less than about 100 mM.
Claim 6
Dependent Claim
27 words
The method of claim 1 wherein said viscosity-increasing agent is selected from at least one of the group consisting of water soluble polymers, sugars, polypeptides, and gelatin.
Claim 7
Dependent Claim
12 words
The method of claim 1 wherein said viscosity-increasing agent is polyvinyl alcohol.
Claim 8
Dependent Claim
20 words
The method of claim 1 wherein said lysate is allowed to stand from about 5 minutes to about 10 minutes.
Claim 9
Dependent Claim
28 words
The method of claim 1 wherein the filter is comprised of a material selected from at least one of the group consisting of cellulose acetate, polyvinylidene, and polycarbonate.
Claim 10
Dependent Claim
18 words
The method of claim 1 wherein the filter comprises cellulose acetate having a pore size of 0.45 microns.
Claim 11
Dependent Claim
12 words
The method of claim 1 wherein the detergent is sodium dodecyl sulfate.
Claim 12
Dependent Claim
13 words
The method of claim 4 wherein said cation concentration is about 10 mM.
Claim 13
Dependent Claim
23 words
The method of claim 1 wherein the step of lysing is performed at a temperature from about 0° C. to about 50° C.
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) | 1996-08-09 | 1997-02-08 | 1997-08-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-08-09 | 2001-02-08 | 2001-08-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-08-09 | 2005-02-08 | 2005-08-10 | $7700.00 | Window Closed / Lapsed |