Process for the purification of DNA on diatomaceous earth

• US 5,075,430 A
• Filed: 12/18/1990
• Issued: 12/24/1991
• Est. Priority Date: 12/12/1988
• Status: Expired due to Term

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Little, Michael C.
• Primary Examiner(s): Rollins, John W.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US07/629,787
• Time in Patent Office: 371 Days
• Field of Search: 935/19, 536/27, 423/335, 435/803
• US Class Current: 536/25.41
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding

Abstract

This invention is directed to a process for the purification of plasmid and other DNA, both single-stranded and double-stranded, by immobilizing the DNA onto diatomaceous earth in the presence of a chaotropic agent and eluting the DNA with water or low salt buffer. The resulting purified DNA is biologically active. Also included in the invention is a process for the immobilization of DNA onto diatomaceous earth in the presence of a chaotropic agent.

Claims

1. In a process for the purification of DNA from a liquid mixture comprising such by:

(a) combining said liquid mixture with a chaotropic agent and contacting the resulting combination mixture with silica by either forming a slurry of said combination mixture with silica, passing said combination mixture through a membrane in which silica is entrapped, or passing said combination mixture through an absorption column packed with silica, to selectively adsorb said DNA onto said silica,

(b) washing said silica with a buffer containing from about 20% to about 95% of a lower alkyl alcohol to remove non-adsorbed matter, and

(c) eluting said DNA from said silica with water or a low salt buffer, the improvement in which said silica is diatomaceous earth.

2. A process in accordance with claim 1 in which said chaotropic agent is a member selected from the group consisting of sodium iodide, sodium perchlorate and sodium trichlorate.

3. A process in accordance with claim 1 in which said lower alkyl alcohol is a member selected from the group consisting of methanol, ethanol and isopropanol.

4. A process in accordance with claim 1 in which said diatomaceous earth has an average particle size of less than about 125 microns.

5. A process in accordance with claim 4 in which said diatomaceous earth has an average particle size of less than about 50 microns.

6. A process in accordance with claim 5 in which said diatomaceous earth has an average particle size of from about 1 micron to about 10 microns.