Stabilized compositions of thermostable DNA polymerase and anionic or zwitterionic detergent
The present invention provides compositions, methods, and kits for protecting thermostable DNA polymerase during amplification reactions conducted at a temperature ranging from about 40° C. to greater than 100° C. The composition comprises a thermostable DNA polymerase and an anionic detergent or zwitterionic detergent.
1. A composition, the composition comprising a thermostable DNA polymerase and an anionic detergent selected from the group consisting of poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether potassium salt; poly(ethylene glycol) monolaurate; polyoxyethylene(150)dinonylphenyl ether; and nonyl nonoxynol-15 phosphate.
2. The composition of claim 1 , wherein the DNA polymerase in the composition retains enzymatic activity for at least 40 days at 25° C.
3. The composition of claim 1 , wherein the DNA polymerase is from a thermophilic bacteria selected from the group consisting of Thermus flavus, Thermus ruber, Thermus thermophilus, Bacillus stearothermophilus, Thermus aquaticus, Thermus lacteus, Thermus rubens , and Methanothermus fervidus.
4. The composition of claim 1 , wherein the DNA polymerase is Taq and the anionic detergent is poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether potassium salt.
5. The composition of claim 1 , wherein the DNA polymerase is Taq and is present at a concentration from about 0.1 unit/μL to about 5 unit/μL; and the detergent is present at a concentration from about 0.001% to about 5% by weight.
6. The composition of claim 1 , further comprising a reducing agent.
7. The composition of claim 1 , further comprising a buffering agent that maintains the composition at a pH of from about 6 to about 9, wherein the buffering agent is selected from the group consisting of MOPS, HEPES, TAPS, Bicine, Tricine, TES, PIPES, MES, or a Tris.
8. The composition of claim 1 , further comprising a mixture of dNTPs selected from the group consisting of dATP, dCTP, dGTP and dTTP.
9. The composition of claim 1 , further comprising a salt selected from the group consisting of potassium chloride, sodium chloride, magnesium chloride, calcium chloride and lithium chloride.
10. The composition of claim 1 , further comprising an agent that increases the efficiency of PCR selected from the group consisting of dimethyl sulfoxide, formamide, and betaine.
11. The composition of claim 1 , wherein the anionic detergent is poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether potassium salt.
12. A kit for amplifying DNA in a thermal cycling process, the kit comprising a thermostable DNA polymerase and an anionic detergent selected from the group consisting of poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether potassium salt, poly(ethylene glycol) monolaurate, polyoxyethylene(150)dinonylphenyl ether, and nonyl nonoxynol-15 phosphate.
13. The kit of claim 12 , wherein the anionic detergent is poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether potassium salt.
14. The kit of claim 12 , further comprising a reducing agent, a buffering agent, and a mixture of dNTPs selected from the group consisting of dATP, dCTP, dGTP, and dTTP.