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- E-ISSN 2288-8985
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Mutation Analysis in β2-Adrenergic Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC)
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2004, v.17 no.1, pp.53-59
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(2004). Mutation Analysis in β2-Adrenergic Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC). Analytical Science and Technology, 17(1), 53-59.
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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 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 β2-adrenergic receptor gene. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.
- keywords
- DHPLC, SSCP, PCR, <TEX>${\beta}_2$</TEX>-adrenergic receptor gene
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