Methods for performing direct enzymatic reactions involving nucleic acid molecules

摘要

Methods for performing a direct enzymatic reaction involving a nucleic acid molecule include performing the enzymatic reaction directly using a biological specimen in a reaction mixture containing a zwitterionic buffer and/or a non-reducing carbohydrate to prevent the biological specimen from inhibiting the enzymatic reaction, in which a nucleic acid molecule present in the biological specimen is not purified before the enzymatic reaction, and obtaining a product from the enzymatic reaction.

主权项

1. A method for preventing a biological specimen from inhibiting a direct enzymatic reaction involving a nucleic acid molecule, the method comprising:
(a) adding a non-reducing carbohydrate to a reaction mixture before contacting the biological specimen to the reaction mixture; wherein the non-reducing carbohydrate is a trehalose and its final concentration in the reaction mixture is at 6% (w/v) to 30% (w/v); (b) contacting the biological specimen to the reaction mixture further comprising a tricine zwitterioninc buffer having a pH 8.3-8.9, wherein the nucleic acid molecule present in the biological specimen is not purified before the enzymatic reaction; and (c) performing the enzymatic reaction directly using the biological specimen in the reaction mixture with preventing the biological specimen from inhibiting the direct enzymatic reaction by the non-reducing carbohydrate.