Literature DB >> 11849627

Real-time PCR for rapid genotyping of angiotensin-converting enzyme insertion/deletion polymorphism.

M H Lin1, C H Tseng, C C Tseng, C H Huang, C K Chong, C P Tseng.   

Abstract

OBJECTIVE: To develop a real-time PCR technique for detection of the insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme (ACE) gene. DESIGN AND METHODS: Three primers were designed for performing real-time PCR in the presence of SYBR Green I as flurochrome followed by melting curve analysis. Forty human genomic DNA that have been genotyped by two-rounds of conventional PCR were used for evaluation of this technique.
RESULTS: Melting curve analysis indicated the melting peak at 73.9 degrees C and 76.2 degrees C corresponding to the presence of I and D alleles, respectively. Comparable genotyping results were obtained by both conventional and real-time PCR. Besides, the mistyping of ID allele individuals by the first run of conventional PCR were accurately genotyped by single-tube real time PCR.
CONCLUSIONS: The real-time PCR method presented in this study provides a rapid and sensitive way for genotyping of ACE gene that may be suitable for large-scale clinical and epidemiologic study.

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Year:  2001        PMID: 11849627     DOI: 10.1016/s0009-9120(01)00281-8

Source DB:  PubMed          Journal:  Clin Biochem        ISSN: 0009-9120            Impact factor:   3.281


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