Nucleic acid amplification using a reversibly inactivated...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/96 (2006.01) C12N 9/12 (2006.01) C12N 9/99 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2184105

The present invention provides a thermostable enzyme which is reversibly inactivated by chemical modification, characterized in that an incubation of said chemically modified thermostable enzyme in an aqueous buffer at alkaline pH at a temperature less than about 25°C results in no significant increase in enzyme activity in less than about 20 minutes, and wherein incubation of said chemically modified enzyme in an aqueous buffer, formulated to about pH 8-9 at 25°C, at a temperature greater than about 50°C results in at least a two-fold increase in enzyme activity in less than about 20 minutes. The chemically modified thermostable enzyme can be used for the amplification of nucleic acids, whereby the activity of the inactivated enzyme is recovered by an incubation of the reaction mixture at an elevated temperature prior to, or as part of, the amplification reaction.

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