Isolation of nucleic acid

• US 8,691,969 B2
• Filed: 06/14/2011
• Issued: 04/08/2014
• Est. Priority Date: 12/12/1994
• Status: Expired due to Fees

• Current Assignee: Life Technologies AS (Perspecta, Inc.)
• Original Assignee: Life Technologies AS (Perspecta, Inc.)
• Inventors: Deggerdal, Arne Helge, Larsen, Frank
• Primary Examiner(s): LEWIS, PATRICK T
• Application Number:
  US13/160,428
• Publication Number:
  US 20110251382A1
• Time in Patent Office: 1,029 Days
• Field of Search: None
• US Class Current: 536/25.4
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

The present invention provides a method of isolating nucleic acid from a sample, said method comprising contacting said sample with a detergent and a solid support, whereby soluble nucleic acid in said sample is bound to the support, and separating said support with bound nucleic acid from the sample. Where the method of the invention is used to isolate DNA, it may conveniently be couple with a further step to isolate RNA from the same sample.

Claims

1. A method of using a particulate solid support for isolating nucleic acid from a sample comprising:

contacting said sample with a detergent and the particulate solid support in the absence of a chaotropic agent,

allowing soluble nucleic acid in said sample to bind directly to the solid support thereby forming a particulate solid support with bound nucleic acid, wherein the binding of the soluble nucleic acid to the solid support is independent of nucleic acid sequence and the binding is non-covalent; and

separating said particulate solid support with bound nucleic acid from the sample, wherein the particulate solid support is not linked to an antibody and wherein the particulate support is not a filter.

2. The method of claim 1, wherein the nucleic acid is DNA.

3. The method of claim 2, wherein the nucleic acid is genomic DNA.

4. The method of claim 1, wherein the nucleic acid is RNA.

5. The method of claim 1, wherein the detergent is anionic.

6. The method of claim 5, wherein the detergent is sodium dodecyl sulphate, or another alkali metal alkylsulphate salt, or sarkosyl.

7. The method of claim 1, wherein the concentration of detergent is 0.2 to 30% (w/v).

8. The method of claim 1, wherein the detergent is contained in a composition additionally comprising one or more monovalent cations, chelating agents or reducing agents.

9. The method of claim 1, wherein the detergent is used in alkaline solution.

10. The method of claim 1, wherein the particulate solid support comprises beads.

11. The method of claim 10, wherein the particulate solid support comprises non-magnetic polymer beads.

12. The method of claim 10, wherein the particulate solid support comprises magnetic beads.

13. The method of claim 1, wherein the solid support has a hydrophobic surface.

14. The method of claim 1, further comprising one or more additional steps to disrupt structural components in the sample or to achieve lysis of cells in the sample.

15. The method of claim 1, wherein the nucleic acid is eluted from the support, following separation from the sample.

16. The method of claim 7, wherein the nucleic acid is eluted by heating.

17. The method of claim 1, wherein the particulate solid support comprises glass or silica.