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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 27, Number 9
BKCSDE 27(9)
September 20, 2006 

 
Title
Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene
Author
Seong Ho Kang*, Sooyoung Jang, Sang-Kyu Park
Keywords
Microchip electrophoresis, Human obesity diagnosis, Single-strand conformation polymorphism, Point mutation
Abstract
We describe an effective method of microchip electrophoresis (ME) based on single strand conformation polymorphism (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.
Page
1346 - 1352
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