Method for reversible modification of thermostable enzymes
AssigneeQiagen GmbH Max-Volmer-Strasse 4
InventorsIgor Ivanov, Dirk Löffert, Jie Kang, Joachim Ribbe, Kerstin Steinert
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Abstract
A method for the amplification of a target nucleic acid is disclosed comprising the steps of reacting a nucleic acid with an amplification reaction mixture and a modified thermostable enzyme, wherein said modified thermostable polymerase is prepared by a reaction of a mixture of a thermostable polymerase and a chemical modifying reagent. The chemical modification reagent is an aldehyde, preferably formaldehyde. Essentially complete inactivation of the enzyme at ambient temperatures is achieved, with recovery of enzymatic activity at temperatures above 50° C.
Claims40
A modified thermostable DNA polymerase, produced by a reaction, under essentially aqueous conditions, of a thermostable DNA polymerase and a modifier reagent at a temperature of less than 50° C., wherein said modifier reagent is an aldehyde of the formula RHC═O, where R is H or an alkyl, aryl, or alkylaldehyde group of 1 to 10 carbon atoms, and wherein said reaction results in a thermally reversible inactivation of the DNA polymerase.
A method for the amplification of a target nucleic acid comprising the step of:
- (a) contacting said nucleic acid with an amplification reaction mixture containing a modified thermostable enzyme, wherein said modified thermostable enzyme is produced by a reaction, under essentially aqueous conditions, of a thermostable enzyme and a modifier reagent at a temperature of less than 50° C., wherein said modifier reagent is an aldehyde of the formula RHC═O, where R is H or an alkyl, aryl, or alkylaldehyde group of 1 to 10 carbon atoms, and wherein said reaction results in thermally reversible inactivation of the enzyme.
Dependencies are inferred from the claim text.