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Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene
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  • Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene
  • Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene
저자명
Kang. Seong-Ho,Jang. Soo-Young,Park. Sang-Kyu
간행물명
Bulletin of the Korean Chemical Society
권/호정보
2006년|27권 9호|pp.1346-1352 (7 pages)
발행정보
대한화학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

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.