Capillary electrophoresis on microchip-based genetic analysis systems are very useful because of rapid analysis, tiny sample/reagent consumption, and portability. Among the many other electrophoretic techniques, capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) analysis is one of promising genotyping method due to its simplicity and versatility. The electrophoretic mobility variation comes from sequence-induced folded conformation difference of single strand DNA. Therefore, any DNA sequence variation theoretically cause shift in the mobility and can be detected upon electrophoresis in non-denaturing condition. However, the most critical limitation of CE-SSCP on microchip is low separation resolution resulting from short separation channel length, thus its application had been limited. To develop high-resolution microchip CE-SSCP analysis system, we investigated the relationship between the resolution of SSCP and the channel width. In this study, we translated CE-SSCP system into a microchip format and improved separation resolution via microchannel width control. A high-resolution microchip CE-SSCP system was demonstrated by using an optimum condition (300μm channel width with PEO-PPO-PEO triblock copolymer). This analysis system was successfully applied to the various genetic analyses including food-borne pathogen detection and single nucleotide polymorphism marker detection.

Development of microchip CE system for single-strand conformation polymorphism analysis

253rd National Meeting (2017)

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