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

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.

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-based non-ionic surfactant selected from the group consisting of polyoxyethylene nonylphenlyether, polyoxyethylene (5) isooctylphenlyther and polyoxyethylene (12) isooctylphenylether, in the range from ≧8% weight/volume to ≦50% 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 component of the polyoxyethylene nonylphenlyether contains from 2 to 150 (CH 2 CH 2 O) units. 
     
     
         4 . The lysis, binding or wash reagent according to  claim 1 , wherein the polyoxyethylene nonylphenlyether is selected from the group consisting of polyoxyethylene(2)nonylphenlyether, polyoxyethylene(5)nonylphenlyether, polyoxyethylene(9)nonylphenlyether, polyoxyethylene(12)nonylphenlyether and polyoxyethylene(100)nonylphenlyether. 
     
     
         5 . The lysis, binding or wash reagent according to  claim 1 , wherein the polyoxyethylene nonylphenlyether is selected from the group consisting of polyoxyethylene(5)nonylphenlyether, polyoxyethylene(9)nonylphenlyether, and polyoxyethylene(12)nonylphenlyether. 
     
     
         6 . The lysis, binding or wash reagent according to  claim 1 , wherein the polyoxyethylene-based non-ionic surfactant is in the range from ≧9% weight/volume to ≦40% weight/volume based on the total volume of the reagent. 
     
     
         7 . The lysis, binding or wash reagent according to  claim 1 , wherein the polyoxyethylene-based non-ionic surfactant is in the range from ≧10% weight/volume to ≦30% weight/volume based on the total volume of the reagent. 
     
     
         8 . The lysis, binding or wash reagent according to  claim 1 , wherein the polyoxyethylene-based non-ionic surfactant is in the range from ≧15% weight/volume to ≦20% weight/volume based on the total volume of the reagent. 
     
     
         9 . The lysis, binding or wash reagent according to  claim 1 , wherein the guanidinium salt is selected from the group consisting guanidinium hydrochloride, guanidinium thiocyanate, guanidinium isothiocyanate, and mixtures of two or more salts thereof. 
     
     
         10 . The lysis, binding or wash reagent according to  claim 1 , wherein the binding reagent further comprises a branched or unbranched alkanol. 
     
     
         11 . The lysis, binding or wash reagent according to  claim 10 , the branched or unbranched alcohol has from one to five carbon atoms. 
     
     
         12 . The lysis, binding or wash reagent according to  claim 11 , 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. 
     
     
         13 . A method for isolating or purifying nucleic acids from a nucleic acid-containing biological sample, comprising the following method steps:
 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,   wherein the lysis composition, the binding composition, or both the lysis composition and the biding composition comprise:
 at least one chaotropic compound that is a guanidinium salt, 
 at least one buffer compound, and 
 at least one polyoxyethylene-based non-ionic surfactant selected from the group consisting of polyoxyethylene nonylphenlyether, polyoxyethylene (5) isooctylphenlyther and polyoxyethylene (12) isooctylphenylether, in the range from ≧0.1% weight/volume to ≦50% weight/volume based on the total volume of the composition. 
   
     
     
         14 . The method of  claim 13 , wherein the binding composition further comprises a branched or unbranched alkanol. 
     
     
         15 . The method of  claim 13 , wherein the polyoxyethylene component of the polyoxyethylene nonylphenlyether contains from 2 to 150 (CH 2 CH 2 O) units. 
     
     
         16 . The method of  claim 15 , wherein the polyoxyethylene nonylphenlyether is selected from the group consisting of polyoxyethylene(2)nonylphenlyether, polyoxyethylene(5)nonylphenlyether, polyoxyethylene(9)nonylphenlyether, polyoxyethylene(12)nonylphenlyether and polyoxyethylene(100)nonylphenlyether. 
     
     
         17 . The method of  claim 13 , wherein the polyoxyethylene nonylphenlyether is selected from the group consisting of polyoxyethylene(5)nonylphenlyether and polyoxyethylene(9)nonylphenlyether and polyoxyethylene(12)nonylphenlyether. 
     
     
         18 . The method according to  claim 13 , wherein the lysis or binding composition comprises a non-ionic surfactant in the range from ≧0.2% weight/volume to ≦30% weight/volume based on the total volume of the composition. 
     
     
         19 . The method of  claim 13 , wherein the lysis or binding composition comprises a non-ionic surfactant in the range from ≧3% weight/volume to ≦10% weight/volume. 
     
     
         20 . The method of  claim 13 , wherein the lysis or binding composition comprises a non-ionic surfactant in the range from ≧3.2% weight/volume to ≦8% weight/volume based on the total volume of the composition. 
     
     
         21 . 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 . 
     
     
         22 . A method for solubilizing lipids of a lipid-containing biological sample, comprising: contacting a lipid-containing biological sample with a polyoxyethylene-based non-ionic surfactant selected from the group consisting of polyoxyethylene nonylphenlyether, polyoxyethylene (5) isooctylphenlyther and polyoxyethylene (12) isooctylphenylether. 
     
     
         23 . A method of preparing the lysis, binding or wash reagent of  claim 1 , comprising: mixing the at least one polyoxyethylene-based non-ionic surfactant with the at least one chaotropic compound and the at least one buffer compound, thereby forming the lysis, bind or washing reagent. 
     
     
         24 . 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,   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 .