High-throughput Single-strand Conformation Polymorphism Analysis of an LPL Gene Mutation by Temperature-controlled On-chip Capillary Electrophoresis

Abstract

Temperature-controlled on-chip capillary electrophoresis (CE) was applied to multi-temperature single-strand conformation polymorphism (SSCP) analysis for the detection of three lipoprotein lipase (LPL) gene mutations. In the SSCP analysis, five different temperature conditions (10, 15, 20, 25, and 30degC) were used to obtain an SSCP pattern. Differences between the electropherograms of Cy5-labeled PCR products of wild/wild homozygote and wild/mutant heterozygote were clearly detected for all four LPL mutations in at least one appropriate temperature condition. These results indicate that our SSCP system is applicable to high-throughput and accurate SSCP screening for locating unknown as well as known single nucleotide polymorphisms (SNPs)