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Genet Test. 2001 Spring;5(1):45-6.

PCR-RFLP genotyping assay for the Bcl I polymorphism of the beta-fibrinogen gene.

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1
Department of Nutrition Sciences, University of Alabama at Birmingham, 35294, USA. sells@uab.edu

Abstract

Polymorphisms of the beta-fibrinogen gene have been shown to affect plasma fibrinogen levels and risk of coronary artery disease (CAD). We were interested in developing an automated, PCR-based genotyping assay for the purpose of exploring relationships between CAD and CAD-associated aortic stiffness and the Bcl I allele of the beta-fibrinogen gene. We have developed a rapid PCR-RFLP assay for the Bcl I polymorphism of the beta-fibrinogen gene. We carried out direct PCR of genomic DNA to facilitate sequencing of the flanking region of the beta-fibrinogen gene. Using this new sequence information, primers were designed which border the site of the Bcl I polymorphism. One of the primers was labeled with a fluorophore to facilitate detection of the fragments. DNA fragment analysis was carried out using an automated capillary electrophoresis instrument (ABI310). We have developed an improved PCR-RFLP high-sample-throughput assay for the semiautomated detection of the Bcl I polymorphism of the beta-fibrinogen gene. This assay will support screening of large sample sizes required for population studies.

PMID:
11336400
DOI:
10.1089/109065701750168707
[Indexed for MEDLINE]
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