Development of the novel high precision low labor load gene-diagnosis system towards realization of tailor-made medical care

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A multiple detection method for single nucleotide polymorphism (SNP) based on the CE separation technique for single base substituted sequence isomers of single stranded DNA was proposed. In the CE separation method where an acidic aqueous urea solution was employed as an electrophoretic buffer solution, the separation of single base substituted sequence isomers of single stranded DNA was achieved according to the difference of the proton dissociation properties of each DNA bases at acidic condition. A clear criterion for determining of existence or nonexistence of SNP by the peak pattern of the electropherogram allows high reliability of the proposed method. The proposed method can be applied to all substitution patterns with identical condition. The proposed method was successfully applied to the SNP detection and genotyping for extracted human gene fragment sample including ALDH2 gene region.