High‐resolution pluronic‐filled microchip CE‐SSCP analysis system via channel width control

Abstract

Although the resolution of CE-SSCP has been significantly improved by using a poly(ethyleneoxide)-poly(propyleneoxide)-poly(ethyleneoxide) (PEO-PPO-PEO; Pluronic(®)) triblock copolymer as a separation medium, CE-SSCP on a microchip format is not widely applicable because their resolution is limited by short channel length. Therefore, a strategy to improve the resolution in channels of limited lengths is highly required for enabling microchip-based CE-SSCP. In this study, we developed a high-resolution CE-SSCP microchip system by controlling the width of the pluronic-filled channel. We tested four different channel widths of 180, 240, 300, and 400 μm, and found that 300 μm showed the highest resolution in the separation of two pathogen specific markers. Potential applications of our method in various genetic analyses were also shown by using SNP markers for spinal muscular atrophy.

Non‐hybridization single nucleotide polymorphism detection and genotyping assay through direct discrimination of single base mutation by capillary electrophoretic separation of single‐stranded DNA

“…Recently, the CE separation of sequential DNA isomers with single-base substitutions and the same chain length has attracted significant attention because it can aid in the identification of gene mutations [5][6][7][8]. Recently, the CE separation of sequential DNA isomers with single-base substitutions and the same chain length has attracted significant attention because it can aid in the identification of gene mutations [5][6][7][8].…”

“…Herein, a new CE-based non-hybridization assay for SNP detection and genotyping was developed. Recently, the CE separation of sequential DNA isomers with single-base substitutions and the same chain length has attracted significant attention because it can aid in the identification of gene mutations [5][6][7][8]. However, it remains a highly challenging task due to its technical difficulties.…”

Separation of single‐base sequential isomers of single‐stranded DNA by capillary electrophoresis and its application in the discrimination of single‐base DNA mutations

“…Past studies employing CE for DNA separation mainly focused on size‐based techniques using molecular‐sieving media such as gel or polymer matrixes . Recently, CE separation of sequential DNA isomers having single‐base substitutions and the same chain length has attracted the attention of researchers because it aids in the identification of gene mutations . However, DNA separation remains a highly challenging task because of a number of technical difficulties.…”