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Mutation Analysis in β2-Adrenergic Receptor Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC)  

Park, Sang-Bum (Dept. of Chemistry, College of Natural Science, Dankook University)
Han, Sang-Man (Dept. of Chemistry, College of Natural Science, Dankook University)
Nam, Youn-Hyoung (Dept. of Chemistry, College of Natural Science, Dankook University)
Jang, Won-Cheoul (Dept. of Chemistry, College of Natural Science, Dankook University)
Publication Information
Analytical Science and Technology / v.17, no.1, 2004 , pp. 53-59 More about this Journal
Abstract
Up to now, methods for the detection of genetic alterations as single strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) have been used. It is too labor-intensive and expensive to serve for routine analysis. Moreover, lower in its sensitivity and specificity being also strongly dependent on the experience of the investigater. To improve these problems, we analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed phase chromatography (IP-RPC). We extracted genomic DNA from 80 asthma patients and then amplified DNA using PCR and analysed PCR product by SSCP and DHPLC. As a result, we analysed mutation frequency is 19 (23.75%) on SSCP and 25 (31.25%) on DHPLC in ${\beta}_2$-adrenergic receptor gene. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.
Keywords
DHPLC; SSCP; PCR; ${\beta}_2$-adrenergic receptor gene;
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