Solubilized phospholipids for stabilizing nucleic acid polymerases
View Patent ↗Compositions and methods are provided that relate to solubilized phospholipids and their use in stabilizing nucleic acid polymerases. For example, a phospholipid with a tail containing at least 8 carbons can be solubilized in the presence of an amphipathic molecule.
1. A composition comprising:
(a) a thermostable polymerase; and
(b) a solubilized phospholipid comprising at least one hydrophobic tail containing 8 to 30 carbons.
2. The composition of claim 1 , wherein the phospholipid is a phosphatidyl lipid.
3. The composition of claim 2 , wherein the phospholipid is a phosphatidylcholine.
4. The composition of claim 1 , wherein the phospholipid is an ether lipid.
5. The composition of claim 1 , wherein the phospholipid is a sphingolipid.
6. The composition of claim 1 , wherein the phospholipid is a diacyl phospholipid.
7. The composition of claim 1 , wherein the phospholipid comprises a plasminogen core.
8. The composition of claim 1 , wherein, in the composition, the phospholipid is solubilized by an amphipathic solubilizer.
9. The composition of claim 8 , wherein the amphipathic solubilizer is a lipid having a hydrophobic tail containing no more than 8 carbons.
10. The composition of claim 1 , wherein the thermostable polymerase is a bacterial polymerase.
11. The composition of claim 1 , wherein the thermostable polymerase is an archael polymerase.
12. The composition of claim 1 , wherein the thermostable polymerase is Taq polymerase or a variant thereof.
13. The composition of claim 1 , further comprising a buffering agent.
14. The composition of claim 13 , wherein the buffering agent is Tris.
15. The composition of claim 1 , further comprising glycerol.
16. The composition of claim 1 , further comprising dNTPs.
17. The composition of claim 1 , wherein the composition comprises a solubilizer and the long-chain phospholipid at a molar ratio in the range of 1:1 to 10:1.
18. A kit comprising:
a thermostable polymerase;
a phospholipid having at least one hydrophobic tail containing 8 to 30 carbons; and
a solubilizer that solubilizes the phospholipid.
19. The kit of claim 18 , wherein the thermostable polymerase, phospholipid and solubilizer are in the same tube.
20. The kit of claim 18 , wherein the thermostable polymerase is in a different tube to the phospholipid and solubilizer.
21. The kit of claim 18 , wherein the phospholipid is a phosphatidyl lipid, a phosphatidylcholine, an ether lipid, a sphingolipid or a diacyl phospholipid, or comprises a plasminogen core.
22. The kit of claim 18 , wherein the stabilizer is a lipid having a hydrophobic tail containing no more than 8 carbons.
23. The kit of claim 18 , wherein the thermostable polymerase is a bacterial polymerase.
24. The kit of claim 18 , wherein the thermostable polymerase is an archael polymerase.
25. The kit of claim 18 , wherein the thermostable polymerase is Taq polymerase or a variant thereof.
26. The kit of claim 18 , further comprising a concentrated reaction buffer.
27. The kit of claim 18 , further comprising dNTPs.