ISOTHERMAL AMPLIFICATION UNDER LOW SALT CONDITION

摘要

Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′ end and a 5′ end, a molecular crowding reagent, and a buffer solution for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.

主权项

1. A kit for amplifying a nucleic acid comprising:
(a) a DNA polymerase having a strand displacement activity; (b) a molecular crowding reagent; and (c) a buffer that provides a final salt concentration between 10 mM to 20 mM during amplification.