Buffers for use with polymerases
Amplification reaction mixtures comprising a cationic surfactant and an anionic surfactant are provided.
1. A solution for storing a polymerase, the solution comprising,
1-20 mM Mg 2+ ;
a polymerase;
an anionic surfactant, wherein the concentration of the anionic surfactant is between 0.0001% and 0.1%; and
a cationic surfactant, wherein the concentration of the cationic surfactant is between 0.0001% and 0.1%.
2. The solution of claim 1 , wherein the anionic surfactant is sodium dodecyl sulfate (SDS).
3. The solution of claim 1 , wherein the cationic surfactant is cetyl trimethylammonium bromide (CTAB).
4. The solution of claim 1 , wherein the anionic surfactant is SDS and the cationic detergent is CTAB.
5. The solution of claim 1 , further comprising a zwitterionic surfactant.
6. The solution of claim 5 , wherein the zwitterionic surfactant is 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS).
7. The solution of claim 5 , wherein the concentration of the zwitterionic surfactant is 0.01-1.0%.
8. The solution of claim 1 , wherein the polymerase is thermostable.
9. A reaction mixture comprising
1-20 mM Mg 2+ ;
a nucleic acid sample;
a polymerase;
an anionic surfactant, wherein the concentration of the anionic surfactant is between 0.0001% and 0.1%; and
a cationic surfactant, wherein the concentration of the cationic surfactant is between 0.0001% and 0.1%.
10. The reaction mixture of claim 9 , further comprising at least one of nucleotides or oligonucleotide primers and a buffer suitable for polymerase chain reaction (PCR).
11. The reaction mixture of claim 9 , wherein the anionic surfactant is sodium dodecyl sulfate (SDS).
12. The reaction mixture of claim 9 , wherein the cationic surfactant is cetyl trimethylammonium bromide (CTAB).
13. The reaction mixture of claim 9 , wherein the anionic surfactant is SDS and the cationic detergent is CTAB.
14. The reaction mixture of claim 9 , further comprising a zwitterionic surfactant.
15. The reaction mixture of claim 14 , wherein the zwitterionic surfactant is 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS).
16. A method of performing a primer extension reaction, the method comprising,
forming a reaction mixture comprising:
a nucleic acid sample that may comprise a target sequence;
a primer that anneals to the target sequence; and
the solution of claim 1 ;
annealing the primer to the target sequence, if present; and
extending the primer by at least one nucleotide with the polymerase.
17. The method of claim 16 , wherein the extending comprises thermocyclic conditions.
18. The method of claim 17 , wherein the extending comprises the polymerase chain reaction (PCR).