PCR-RFLP genotyping assay for the Bcl I polymorphism of the beta-fibrinogen gene.
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.
Sell
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SM
Department of Nutrition Sciences, University of Alabama at Birmingham, 35294, USA. sells@uab.edu
Song
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C
Booyse
F M
FM
D000483:Alleles; D001331:Automation; D003327:Coronary Disease / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D005340:Fibrinogen / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013997:Time Factors
D005340:Fibrinogen; C041666:endodeoxyribonuclease BclI; D015252:Deoxyribonucleases, Type II Site-Specific