Method for activating a nucleic acid for a polymerase reaction
First Claim
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1. A method for activating a non-damaged nucleic acid for a strand displacement reaction with the steps:
- (a) heating a reaction mixture comprising a non-damaged nucleic acid to a temperature between 55°
C. to 80°
C.,(b) cooling the reaction mixture to a temperature of 4°
C. to 45°
C., and(c) adding a polymerase to the cooled reaction mixture of step (b),wherein the polymerase added in step (c) starts the strand displacement reaction,wherein the reaction mixture is not an alkali solution, andwherein two or more primers are added prior to or subsequent to step (a).
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Abstract
The present invention concerns a method for activating a nucleic acid for a polymerase reaction with the steps: (a) Heating a nucleic acid to a temperature of 55° C. to 80° C., (b) cooling the nucleic acid to a temperature at which a polymerase shows no substantial decrease in activity, and (c) starting the polymerase reaction by the addition of a heat-labile polymerase to the nucleic acid.
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Citations
14 Claims
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1. A method for activating a non-damaged nucleic acid for a strand displacement reaction with the steps:
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(a) heating a reaction mixture comprising a non-damaged nucleic acid to a temperature between 55°
C. to 80°
C.,(b) cooling the reaction mixture to a temperature of 4°
C. to 45°
C., and(c) adding a polymerase to the cooled reaction mixture of step (b), wherein the polymerase added in step (c) starts the strand displacement reaction, wherein the reaction mixture is not an alkali solution, and wherein two or more primers are added prior to or subsequent to step (a). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for activating a non-damaged nucleic acid for a strand displacement reaction with the steps:
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(a) heating a reaction mixture comprising a non-damaged nucleic acid to a temperature between 55°
C. to 65°
C.,(b) cooling the reaction mixture to a temperature of 4°
C. to 45°
C., and(c) adding a polymerase to the cooled reaction mixture of step (b), wherein the polymerase added in step (c) starts the strand displacement reaction, and wherein two or more primers are added prior to or subsequent to step (a).
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Specification