Device and process for isolating nucleic acids from cell suspension

• US 5,652,141 A
• Filed: 05/16/1995
• Issued: 07/29/1997
• Est. Priority Date: 10/26/1990
• Status: Expired due to Term

• Current Assignee: Oiagen GmbH
• Original Assignee: Oiagen GmbH
• Inventors: Henco, Karsten, Colpan, Metin, Feuser, Petra
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): PRATS, FRANCISCO CHANDLER
• Application Number:
  US08/442,100
• Time in Patent Office: 805 Days
• Field of Search: 435/6, 435/270, 435/288, 435/259, 536/127
• US Class Current: 435/270
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

A device and a process for isolating nucleic acids by lysing intact cells and removing nucleic acids emerging from the lysed cells by the following steps:

a) the cells are immobilized in a porous matrix, with the size of matrix voids being in the range of the type of cell to be lysed;

b) the cells are lysed;

c) the nucleic acids are fixated on the matrix surface, and subsequently

d) are eluted.

Claims

1. A process for isolating nucleic acids by lysing blood cells, substantially all of which are intact, and removing the nucleic acids emerging from the lysed cells, comprising the steps of:

a) immobilizing the cells in a porous matrix having ion exchange properties and having voids each having a size sufficient to capture one of the cells;

b) lysing the cells;

c) fixing the emerging nucleic acids on the matrix surface by ion exchange, and subsequently;

d) eluting the nucleic acids.

2. The process according to claim 1, wherein the matrix has a void size of from 1 to 50 μ

m.

3. The process according to claim 2, wherein the void size is from 5 to 15 μ

m.

4. The process according to claim 1, wherein the porous matrix comprises particles each having a particle size of 10 to 50 μ

m.

5. The process according to claim 4, wherein the particle size is from 15 to 25 μ

m.

6. The process according to claim 1, wherein the cell lysing is effected by ultrasonic waves, shear forces, detergents, enzymes, osmotic shock, or combinations thereof.

7. The process according to claim 1, wherein the nucleic acids are eluted by a buffer or by alkaline elution.

8. The process according to claim 1, wherein said matrix is washed between at least one of steps a) and b), and b) and c), and c) and d).

9. The process according to claim 1, wherein the matrix is an inorganic or organic polymer.

10. The process according to claim 9, wherein the matrix has a void size of from 1 to 50 μ

m.

11. The process according to claim 10, wherein the void size is from 5 to 15 μ

m.

12. In a process for isolating nucleic acids by lysing blood cells and removing the nucleic acids emerging from the lysed cells, the improvement comprising the steps of:

a) before lysing, immobilizing the blood cells in a porous matrix having ion exchange properties and having voids each having a size sufficient to capture one of the cells;

b) lysing the captured cells;

c) fixing the emerging nucleic acids on the matrix surface by ion exchange, and subsequently;

d) eluting the nucleic acids.