title: Detection of specific alleles by using allele-specific primer extension followed by capture on solid support.
abstract: This article describes a method for determining whether a particular nucleic acid sequence is present in a sample and for discriminating between any two nucleic acid sequences if such sequences differ only by a single nucleotide. The method entails extension of a novel two-component primer on templates that may or may not include a target nucleic acid sequence. The 3' portion of the primer is complementary to a portion of the template adjacent to the target sequence (for example, the polymorphic nucleotide). The 5' portion of the primer is complementary to a different preselected nucleic acid sequence. Extension of the 3' portion of the primer with a labeled deoxynucleoside triphosphate yields a labeled extension product, but only if the template includes the target sequence. The presence of such a labeled primer-extension product is detected by hybridization of the 5' portion to the preselected sequence. The preselected sequence is immobilized on a solid support. The method has been applied to genotyping individuals for the two-allele polymorphism of the human tyrosinase gene.
authors: Department of Molecular Biochemistry, Beckman Research Institute of the City of Hope, Duarte, California.
authors: L
authors: L
authors: Ugozzoli
authors: J M
authors: JM
authors: Wahlqvist
authors: A
authors: A
authors: Ehsani
authors: B E
authors: BE
authors: Kaplan
authors: R B
authors: RB
authors: Wallace
mesh_terms: D000483:Alleles; D001483:Base Sequence; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000235:genetics; D011110:Polymorphism, Genetic
chemical_list: D004247:DNA; D014442:Monophenol Monooxygenase