Lysis, binding and/or wash reagent for isolating and/or purifying nucleic acids

Patent metadata

Field	Value
Publication number	US-9617532-B2
Application number	US-99430809-A
Country	US
Kind code	B2
Filing date	May 25, 2009
Priority date	May 30, 2008
Publication date	Apr 11, 2017
Grant date	Apr 11, 2017

Abstract

The present invention relates to a lysis, binding and/or wash reagent for isolating and/or purifying nucleic acids and a method for isolating and/or purifying nucleic acids.

First claim

The invention claimed is:

1. A lysis, binding or wash reagent comprising:
at least one chaotropic compound that is a guanidinium salt,
at least one buffer compound, and
at least one polyoxyethylene fatty alcohol ether selected from the group consisting of polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, and polyoxyethylene oleyl ether in the range from ≧8% weight/volume to ≦30% weight/volume based on the total volume of the reagent.

2. The lysis, binding or wash reagent of claim 1 , wherein the at least one buffer compound is selected from the group consisting of tris(hydroxymethyl)aminomethane, N-(tri(hydroxymethyl)methyl)glycine, N,N-bis(2-hydroxyethyl)glycine, 3-(N-morpholino)propanesulphonic acid, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulphonic acid), piperazine-1,4-bis(2-ethanesulphonic acid), N-cyclohexyl-2-aminoethanesulphonic acid, 2-(N-morpholino)ethanesulphonic acid, and phosphate.

3. The lysis, binding or wash reagent according to claim 1 , wherein the polyoxyethylene fatty alcohol ether comprises a polyoxyethylene component containing from 2 to 150 (CH 2 CH 2 O) units.

4. The lysis, binding or wash reagent according to claim 3 , wherein the polyoxyethylene fatty alcohol ether is polyoxyethylene cetyl ether.

5. The lysis, binding or wash reagent according to claim 1 , wherein the polyoxyethylene fatty alcohol ether is in the range from ≧10% weight/volume to ≦30% weight/volume based on the total volume of the reagent.

6. The lysis, binding or wash reagent according to claim 1 , wherein the polyoxyethylene fatty alcohol ether is in the range from ≧15% weight/volume to ≦20% weight/volume based on the total volume of the reagent.

7. The lysis, binding or wash reagent according to claim 1 , wherein the guanidinium salt is selected from the group consisting of guanidinium hydrochloride, guanidinium thiocyanate, guanidinium isothiocyanate, and mixtures of two or more salts thereof.

8. The lysis, binding or wash reagent according to claim 1 , wherein the lysis, binding or wash reagent is a binding reagent and further comprises a branched or unbranched alcohol.

9. The lysis, binding or wash reagent according to claim 8 , wherein the branched or unbranched alcohol has from one to five carbon atoms.

10. The lysis, binding or wash reagent according to claim 9 , wherein the branched or unbranched alcohol is selected from the group consisting of methanol, ethanol, isopropanol, n-propanol, n-butanol, branched or unbranched butanol or pentanol, and mixtures thereof.

11. A method for isolating or purifying nucleic acids from a nucleic acid-containing biological sample, comprising:
a) lysing the biological sample in the presence of a lysis composition,
b) immobilizing the released nucleic acid(s) on a matrix based on one or more silicon oxide compound(s) in the presence of a binding composition,
c) optionally washing the nucleic acid(s) immobilized on the matrix, and
d) optionally removing the bound nucleic acid(s),
wherein the lysis composition, the binding composition, or both the lysis composition and the binding composition comprise:
at least one chaotropic compound that is a guanidinium salt,
at least one buffer compound, and
at least one polyoxyethylene fatty alcohol ether selected from the group consisting of polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, and polyoxyethylene oleyl ether in the range from ≧8% weight/volume to ≦30% weight/volume based on the total volume of the composition.

12. The method of claim 11 , wherein the binding composition further comprises a branched or unbranched alcohol.

13. The method of claim 11 , wherein the polyoxyethylene fatty alcohol ether comprises a polyoxyethylene component containing from 2 to 150 (CH 2 CH 2 O) units.

14. The method of claim 13 , wherein the polyoxyethylene fatty alcohol ether is polyoxyethylene cetyl ether.

15. A kit for isolating or purifying nucleic acids from a nucleic acid-containing biological sample, comprising a lysis, binding or wash reagent of claim 1 .

16. A method of preparing a storage-stable binding, lysis or wash reagent, comprising:
mixing at least one polyoxyethylene fatty alcohol ether with a chaotropic compound and a buffer compound, wherein
the chaotropic compound is a guanidinium salt,
the at least one polyoxyethylene fatty alcohol ether is selected from the group consisting of polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, and polyoxyethylene oleyl ether, and
the at least one polyoxyethylene fatty alcohol ether is in the range from ≧8% weight/volume to ≦30% weight/volume based on the total volume of the reagent.

17. The method of claim 16 , wherein the polyoxyethylene fatty alcohol ether is polyoxyethylene cetyl ether.

18. A method for isolating or purifying nucleic acids from a nucleic acid-containing biological sample, comprising:
(a) lysing the biological sample using a lysis reagent,
(b) immobilizing the released nucleic acid(s) on a matrix based on one or more silicon oxide compound(s) using a binding reagent,
(c) optionally washing the nucleic acid(s) immobilized on the matrix using a wash reagent, and
(d) optionally removing the bound nucleic acid(s),
wherein one, two, or three of the lysis reagent, the binding reagent, and the wash reagent are the lysis, binding or wash reagent according to claim 1 .

Assignees

• Fabis Roland
• Homann-Wischinski Anke
• Voss Thorsten
• Hanselle Thomas
• Qiagen Gmbh

Inventors

• Fabis Roland
• Homann-Wischinski Anke
• Voss Thorsten
• Hanselle Thomas

Classifications

• C12N1/06

  Lysis of microorganisms · CPC title

• C12N15/1003 Primary

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• C12N15/1006 Primary

  by means of a solid support carrier, e.g. particles, polymers · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12N15/1013

  by using magnetic beads · CPC title