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Genotyping of the 19-bp insertion/deletion polymorphism in the 5′ flank of β-hydroxylase gene by dissociation analysis of allele-specific PCR products

Genotyping of the 19-bp insertion/deletion polymorphism in the 5′ flank of β-hydroxylase gene by... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Clinical Chemistry and Laboratory Medicine (CCLM) de Gruyter

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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Publisher
de Gruyter
Copyright
Copyright © 2005 by the
ISSN
1434-6621
eISSN
1437-4331
DOI
10.1515/CCLM.2005.154
pmid
16176167
Publisher site
See Article on Publisher Site

Abstract

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.

Journal

Clinical Chemistry and Laboratory Medicine (CCLM) – de Gruyter

Published: Sep 1, 2005

Keywords: allele drop-out; dissociation analysis; dopamine β-hydroxylase gene; enzymatic amplification; genotyping; insertion-deletion polymorphism

References

  • A haplotype at the DBH locus , associated with low plasma dopamine beta - hydroxylase activity , also associates with cocaine - induced paranoia
    Cubells
  • Dopamine beta - hydroxylase gene and schizophrenia phenotypic variability : A genetic association study
    Yamamoto
  • Mistyping frequency of the angiotensin - converting enzyme gene polymorphism and an improved method for its avoidance
    Odawara
  • Preferential PCR amplification of alleles : mechanisms and solutions
    Walsh
  • Serum dopamine β - hydroxylase
    Weinshilboum
  • Melting curve SNP genotyping : a useful approach for diallelic genotyping in forensic science
    Ye
  • A quantitative - trait analysis of human plasma - dopamine beta - hydroxylase activity : evidence for a major functional polymorphism at the DBH locus
    Zabetian
  • Mistyping of the human angiotensin - converting enzyme gene polymorphism : frequency , causes and possible methods to avoid errors in typing
    Ueda
  • Elevated sympathetic nerve activity in borderline hypertensive humans : evidence from direct intraneural recordings
    Anderson
  • A general method for the isolation of RNA complementary to DNA
    Bolton
  • Genetically determined low maternal serum dopamine beta - hydroxylase levels and the etiology of autism spectrum disorders
    Robinson
  • Human genetics of plasma dopamine beta - hydroxylase activity : applications to researchin psychiatry and neurology
    Cubells
  • Validity of PCR with emphasis on variable number of tandem repeat analysis
    Kaiser