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Abstract |
Rapid Detection of the R389G Polymorphism of ADRB1 by Real Time PCR Using Fluorescence Resonance Energy Transfer. |
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The β1-adrenoceptor plays a crucial role in the regulation of myocardial contractility and heart rate. A functionally important polymorphism at codon 389 has been associated with heart failure and hypertension. The conventional method of detecting this polymorphism is a PCR-assay followed by digestion with specific restriction enzyme (PCR-RFLP) polymorphism. We aimed to establish an alternative, fast genotyping method by real time PCR, which combines high speed DNA-amplification and real time fluorescence monitoring using fluorescence resonance energy transfer. Venous blood of 50 healthy volunteers was examined for the Arg389Gly polymorphism using both methods. The results from PCR-RFLP were compared to those obtained with real time fluorescence PCR and showed a 100% correlation. DNA sequencing confirmed a 100% concordance. This alternative technique is significantly less time consuming than traditional PCR-RFLP and facilitates the analysis of a large numbers of samples. |
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