Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene

  • Kang, Seong-Ho (Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University) ;
  • Jang, Soo-Young (Korean Minjok Leadership Academy) ;
  • Park, Sang-Kyu (Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University)
  • Published : 2006.09.20
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Abstract

We describe an effective method of microchip electrophoresis (ME) based on single strand conformation poly-morphism (SSCP) analysis to rapidly detect the point mutation, Leu72Met, in a human obesity gene. The 207-bp dsDNA in the Leu72Met region, an estimate of the child obesity DNA mutant, was amplified by polymerase chain reaction (PCR) and submitted to a conventional glass microchip analysis with a sieving matrix of 1.75% poly(vinylpyrrolidone) (Mr 1 300 000), 1.0% poly(ethyleneoxide) (Mr 600 000) and 5.0% w/w glycerol. When combined with base stacking (BS) with hydroxide ions, the SSCP-ME provided rapid analysis as well as sensitive detection. The detection sensitivity was effectively enhanced in the OH- concentration range of 0.01-0.025 M NaOH. The sensitivity and speed of this ME-based SSCP methodology for the rapid detection of Leu72Met point mutations makes this an attractive method for diagnosing childhood obesity in a clinical diagnostic laboratory.

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