title: Direct molecular haplotyping by melting curve analysis of hybridization probes: beta 2-adrenergic receptor haplotypes as an example.
abstract: Direct determination of the association of multiple genetic polymorphisms, or haplotyping, in individual samples is challenging because of chromosome diploidy. Here, we describe the ability of hybridization probes, commonly used as genotyping tools, to establish single nucleotide polymorphism (SNP) haplotypes in a single step. Three haplotypes found in the beta 2-adrenergic receptor (beta2AR) gene and characterized by three different SNPs combinations are presented as examples. Each combination of SNPs has a unique stability, recorded by its melting temperature, even when intervening sequences from the template must loop out during probe hybridization. In the course of this study, two haplotypes in beta2AR not described previously were discovered. This approach provides a tool for molecular haplotyping that should prove useful in clinical molecular genetics diagnostics and pharmacogenetic research where methods for direct haplotyping are needed.
authors.affiliation: Institute for Clinical and Experimental Pathology, ARUP Laboratories 500 Chipeta Way, Salt Lake City, UT 84108, USA. pontkig@aruplab.com
authors.forename: Genevieve
authors.initials: G
authors.lastname: Pont-Kingdon
authors.forename: Elaine
authors.initials: E
authors.lastname: Lyon
mesh_terms: D001483:Base Sequence; D015342:DNA Probes; D031541:Fluorescence Resonance Energy Transfer; D006239:Haplotypes; D006358:Hot Temperature; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics*; D015203:Reproducibility of Results
chemical_list: D015342:DNA Probes; D018343:Receptors, Adrenergic, beta-2