Reaction mixtures
The present invention provides stable dried reaction mixtures, methods for their preparation, methods for their use, and kits comprising them. The stable dried reaction mixtures are useful in many recombinant DNA techniques, especially nucleic acid amplification by the polymerase chain reaction (PCR).
1. A method of preparing a dry reaction mixture composition, the method comprising drying a reaction mixture in aqueous form by air drying at 45° C. in the absence of lyophilization or vacuum-drying, wherein the reaction mixture in aqueous form comprises at least one nucleic acid amplification-related enzyme, nucleotide triphosphates, manganese acetate (Mn(OAc) 2 ), and sucrose.
2. The method of claim 1 , wherein the dry reaction mixture retains activity upon storage under conditions that are, or are equivalent to, 45° C. for 3 months.
3. The method of claim 1 , wherein sucrose is at a concentration between about 50 mM and about 1000 mM in the reaction mixture in aqueous form.
4. The method of claim 1 , wherein the at least one nucleic acid amplification-related enzyme is a thermostable polymerase selected from the group consisting of Thermus aquaticus Taq DNA polymerase, Thermus sp. Z05 polymerase, Thermus sp. Z05-D polymerase, Thermus flavus polymerase, Thermotoga maritima polymerase, TMA-25 polymerase, TMA-30 polymerase, Tth DNA polymerase, as well as modified thermostable polymerases, or any combination thereof.
5. The method of claim 4 wherein the at least one nucleic acid amplification-related enzyme is a thermostable polymerase selected from Thermus sp. Z05 polymerase or Thermus sp. Z05-D polymerase.