Genotyping of the 19-bp insertion/deletion polymorphism in the 5′ flank of β-hydroxylase gene by dissociation analysis of allele-specific PCR products
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Henrik Berg Rasmussen
Abstract
The 19-bp insertion/deletion polymorphism in the 5′ flank of the dopamine β-hydroxylase ( DBH) gene has been associated with psychiatric disorders. We have developed a simple, reliable and inexpensive closed-tube assay for genotyping of this polymorphism based upon T m determination of amplified DNA fragments. Mistyping of heterozygote samples due to preferential allele amplification was prevented by use of an optimized concentration of Mg 2+, addition of dimethyl sulfoxide and annealing/extension at an appropriate temperature. Comparison of results achieved by the closed-tube assay and a conventional approach based upon agarose gel electrophoresis of amplified fragments revealed complete concordance between the two procedures. The insights obtained in this study may be utilized to develop assays based upon dissociation analysis of PCR products for genotyping of other insertion/deletion polymorphisms.
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©2005 by Walter de Gruyter Berlin New York
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- The future of clinical laboratories: more testing or knowledge services?
- Apolipoprotein A5 and triglyceridemia. Focus on the effects of the common variants
- Genotyping of the 19-bp insertion/deletion polymorphism in the 5′ flank of β-hydroxylase gene by dissociation analysis of allele-specific PCR products
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- Systemic levels of interleukin-6 and matrix metalloproteinase-9 in patients with multiple myeloma may be useful as prognostic indexes of bone disease
- Analysis of the applicability of artificial neural networks for studying blood plasma: determination of magnesium ion concentration as a case study
- The effect of long-term storage on measured plasma lactate concentrations and prospective lactate results from a multicenter trial of antiretroviral therapy
- Evaluation of renal function in intensive care: plasma cystatin C vs. creatinine and derived glomerular filtration rate estimates
- Multicentre physiological reference values for some urinary component-to-creatinine (creatininium) concentration ratios
- Diagnostic and analytical performance of a screening panel for allergy
- Preliminary evaluation of the Iris IQ™ 200 automated urine analyser
- The Asp298 but not the C-786 genotype of the endothelial nitric oxide synthase is reduced with age in healthy Swiss men
- Identification of preanalytical mistakes in the stat section of the clinical laboratory
- Congress of Clinical Chemistry and Laboratory Medicine 2nd Annual Conference of the German United Society for Clinical Chemistry and Laboratory Medicine (DGKL) together with the Austrian Society for Laboratory Medicine and Clinical Chemistry (ÖGLMKC) Jena, Germany, October 6–8, 2005
- SIBioC 2005 37th National Congress of the Italian Society of Clinical Biochemistry and Clinical Molecular Biology Rome, Italy, October 11–14, 2005
