| 摘要 |
PURPOSE: To optimize the amounts of a specimen and a reagent and to enable the preparation and analysis of a small amount of a specimen in high efficiency by setting the objective DNA concentration, the concentration of a fluorescence-labeled DNA primer to be hybridized to the objective DNA, the concentration of a complemental strand synthetase and the amount of the enzyme at respective specific levels. CONSTITUTION: The preparation and analysis of the objective DNA are carried out by separating the objective DNA 1 to be used as a template into a single chain template DNA+ chain 2 and a single chain template DNA-chain by alkali denaturation or heat-shock in the presence of a fluorescence-labeled DNA primer 4, heating the chains at 37 deg.C for 20min to hybridize the primer 4 to the single chain template DNA+ chain 2 and synthesizing a complementary strand from the hybridized primer 5 by the action of a complementary strand synthetase to prepare a slight amount of the objective DNA specimen. In the above process, the mode of hybridization is formulated, a reaction rate constant is determined from the experimental data by using the formulated mode, the amounts of the specimen and reagent are optimized and the reaction is carried out at the objective DNA concentration of 10<-8> M, the fluorescence-labeled DNA primer concentration of 10<-7> to 10<-8> M, the complementary strand synthetase concentration of <=0.1 unit and the enzyme amount of <=5μL.
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