Stable lysis buffer mixture for extracting nucleic acids

Abstract

The invention relates to a lysis buffer mixture that is stable in storage for isolating nucleic acids from biological, preferably diagnostic samples. The mixture is preferably associated with an extraction control. The aim of the invention is to provide an improved nucleic acid extraction system, which is cost-effective, stable and easy to use, thus fulfilling the requirements of a modern nucleic acid extraction system and containing, among other things, extraction controls. The invention relates to a lysis buffer mixture for isolating nucleic acids, said mixture containing non chaotropic salts, a special selection of detergents, a defined quantity of at least one nucleic acid as an extraction control, optionally lytic enzymes, optionally carrier nucleic acids and optionally other additives.

1 . Storage-stable, freeze-dried lysis buffer mixture, manufactured by all the components of the lysis buffer mixture existing together in an aqueous solution and being freeze-dried, for the isolation of nucleic acids from arbitrary complex initial substances, comprising:
 non-chaotropic salts;   at least one detergent;   a defined quantity of at least one nucleic acid as an extraction control;   optionally lytic enzymes; and   optionally carrier nucleic acids.   
     
     
         2 . The lysis buffer mixture of  claim 1 , wherein the non-chaotropic salts contain cations selected from the group consisting of monovalent cations, bivalent cations, multivalent cations, and mixtures thereof. 
     
     
         3 . The lysis buffer mixture of  claim 2  wherein the cations are selected from the group consisting of ammonium ions, sodium ions, potassium ions, magnesium ions, calcium ions, zinc ions and manganese ions. 
     
     
         4 . The lysis buffer mixture of  claim 1 , wherein the lysis buffer mixture has a pH value between 5 and 9. 
     
     
         5 . The lysis buffer mixture of  claim 1  wherein the at least one detergent is selected from the group consisting of cetyltrimethylammonium bromide, Tween 20, Triton X-100, and sodium dodecyl sulfate. 
     
     
         6 . The lysis buffer mixture of  claim 1 , wherein the at least one detergent is cetyltrimethylammonium bromide. 
     
     
         7 . The lysis buffer mixture of  claim 1 , wherein the at least one detergent is present in the range of 0.1-5% of the total quantity. 
     
     
         8 . The lysis buffer mixture of  claim 1 , wherein cetyltrimethylammonium bromide (CTAB) is present as 2% of the total quantity. 
     
     
         9 . The lysis buffer mixture of  claim 1 , wherein the defined quantity of at least one nucleic acid as an extraction control is selected from the group consisting of a synthetic nucleic acid, a nucleic acid manufactured via molecular biology methods, and a native nucleic acid. 
     
     
         10 . The lysis buffer mixture of  claim 1 , wherein the defined quantity of at least one nucleic acid for extraction control is selected from the group consisting of at least one DNA, at least one RNA, and mixtures of at least one DNA and at least one RNA. 
     
     
         11 . (canceled) 
     
     
         12 . The lysis buffer mixture of  claim 1 , comprising lytic enzymes, wherein the lytic enzymes are proteinases. 
     
     
         13 . The lysis buffer mixture of  claim 1 , comprising lytic enzymes, wherein the lytic enzymes are proteinase K. 
     
     
         14 . The lysis buffer mixture of  claim 1 , comprising lytic enzymes, wherein the lytic enzymes are proteinase K and lysozyme. 
     
     
         15 . The lysis buffer mixture of  claim 1 , comprising carrier nucleic acids, wherein the carrier nucleic acids are synthetic nucleic acids or isolates from biological materials. 
     
     
         16 . The lysis buffer mixture of  claim 1 , comprising carrier nucleic acids, wherein the carrier nucleic acids are poly A RNA, tRNA, salmon sperm DNA, or herring sperm DNA. 
     
     
         17 . The lysis buffer mixture of  claim 1 , wherein the lysis buffer mixture is a solid, storage-stable formulation in ready-to-use reaction vessels. 
     
     
         18 . The lysis buffer mixture of  claim 17 , wherein the ready-to-use reaction vessels are 96 well microtitre plates, 384 well microtitre plates, or 0.5-5 ml reaction vessels. 
     
     
         19 . The lysis buffer mixture of  claim 17 , wherein the ready-to-use reaction vessels are 2 ml reaction vessels. 
     
     
         20 . The lysis buffer mixture of  claim 1 , wherein the lysis buffer mixture is in a tablet form. 
     
     
         21 . A method for isolation of nucleic acids from arbitrary complex initial substances comprising using the lysis buffer mixture of  claim 1 . 
     
     
         22 . The method of  claim 21 , wherein the nucleic acids to be isolated are DNA, RNA, or DNA in combination with RNA, and the arbitrary complex initial substances are microorganisms. 
     
     
         23 . A reaction kit for an extraction of nucleic acids comprising the lysis buffer mixture of  claim 1 . 
     
     
         24 . A reaction kit for an extraction of nucleic acids comprising lysis vessels with the lysis buffer mixture of  claim 1 , a binding buffer on an alcohol basis, an elution buffer and a solid phase for a binding of the nucleic acids. 
     
     
         25 . The reaction kit of  claim 24 , further comprising at least one washing buffer. 
     
     
         26 . The reaction kit of  claim 25 , for an extraction of virus nucleic acids or for an extraction of bacterial nucleic acids from diagnostic samples. 
     
     
         27 . A reaction kit for an extraction of nucleic acids, comprising entailing at least one 2 ml lysis vessel with the lysis buffer mixture of  claim 1 , a binding buffer on an isopropyl alcohol basis, a first washing buffer, a second washing buffer, an elution buffer, and spin filters as absorption phase. 
     
     
         28 . (canceled)