Methods and compositions for amplification of nucleic acids

摘要

The present invention provides methods, compositions, and kits for storing and enhancing the activity of polymerases and particularly thermostable polymerases. The methods comprise mixing a thermostable polymerase with at least one zwitterionic or ylide surfactant that has at least one PEO group. In another aspect the polymerase is mixed with a blocker such as PLURONIC® or TETRONIC® or an amine N-oxide derivative thereof. The thermostable polymerase may be reversibly inactivated by treatment with 2-(Methylsulfonyl)ethyl 4-nitrophenyl carbonate. Compositions and kits for performing the process according to the invention are also provided.

主权项

1. A composition comprising a thermostable polymerase that has been reversibly modified with a polymerase-modifying reagent of the following structure: wherein Y=N-hydroxysuccinimide (NHS), p-nitrophenol (pNP) or imidazole;
X=CONMe2, MeSO2, CN, NO2, CO2Me, (CH2)2SMe, N(CH3)CHO, or CH2C(O)CH3;and a zwitterionic amine oxide.