Process for isolating nucleic acid

• US 5,234,809 A
• Filed: 07/01/1991
• Issued: 08/10/1993
• Est. Priority Date: 03/23/1989
• Status: Expired due to Term

• Current Assignee: AKZO NV (Nouryon BV)
• Original Assignee: AKZO NV (Nouryon BV)
• Inventors: Kievits, Tim, Boom, Willem R., Adriaanse, Henriette M. A., Lens, Peter F.
• Primary Examiner(s): Nucker, Christine M.
• Assistant Examiner(s): STUCKER, JEFFREY J
• Application Number:
  US07/728,007
• Time in Patent Office: 771 Days
• Field of Search: 435/6, 435/91, 422/61, 536/27
• US Class Current: 435/91.2
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12N 15/1017 by filtration, e.g. using filters, frits, membranes
• 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: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C12Q 2563/155 Particles of a defined size, e.g. nanoparticles

Abstract

The invention relates to a process, a combination of means for isolating nucleic acid from a nucleic acid-containing starting material and a testkit in order to amplify the nucleic acid obtained by said process. More in particular, the invention relates to a process and a kit for isolating nucleic acid from a nucleic acid-containing biological material such as whole blood, blood serum, urine, feces, cell cultures and the like.

Claims

1. A process for isolating nucleic acid from a nucleic acid-containing starting material comprising mixing the starting material, a chaotropic substance and a nucleic acid binding solid phase, separating the solid phase with the nucleic acid bound thereto from the liquid, and washing the solid phase nucleic acid complexes.

2. A process according to claim 1, wherein the starting material employed is a nucleic acid-containing biological material.

3. A process according to claim 1, wherein the chaotropic substance is at least one compound selected from the group consisting of a guanidinium salt, sodium iodide, potassium iodide, sodium thiocyanate and urea.

4. A process according to claim 3, wherein the guanidinium salt employed is guanidinium (iso)thiocyanate.

5. A process according to claim 1, wherein the nucleic acid binding solid phase is selected from the group consisting of silica particles, cuvettes, microtiter plates, filter membranes, polystyrene beads and nitrocellulose paper.

6. A process according to claim 1 for isolating DNA or RNA.

7. A process according to claim 1, wherein the solid phase comprises silica particles having a particle size ranging substantially between 0.05 and 500 μ

m.

8. A process according to claim 1, wherein the solid phase comprises silica particles having a particle size ranging substantially between 0.1 and 200 μ

m.

9. A process according to claim 1, wherein the solid phase comprises silica particles having a particle size ranging substantially between 1 and 200 μ

m.

10. A process according to claim 1, wherein the solid phase-nucleic acid complexes are separated from the liquid by sedimentation and wherein the complexes are washed with a chaotropic substance-containing washing buffer.

11. A process according to claim 10, wherein the solid phase-nucleic acid complexes washed with washing buffer are further washed successively with one or more alcohol-water solutions and then dried.

12. A process according to claim 11, wherein the nucleic acid present in the washed and dried solid phase-nucleic acid complexes is eluted by means of an aqueous buffer suitable for eluting nucleic acid.

13. A process to amplify captured nucleic acids comprising mixing the starting material, a chaotropic substance and a nucleic acid binding solid phase, separating the solid phase with the nucleic acid bound thereto from the liquid, washing the solid phase nucleic acid complexes and contacting the solid phase-nucleic acid complexes with a mixture comprising components that amplify the nucleic acid.

14. A test kit for carrying out the process according to claim 13 comprising components to amplify nucleic acid.