title: Rapid classification of phenylketonuria genotypes by analysis of heteroduplexes generated by PCR-amplifiable synthetic DNA.
abstract: We describe a rapid and simple method for phenylketonuria genotyping which identifies five point mutations within exon 12 of the human phenylalanine hydroxylase gene. The method involves PCR amplification of the target exon and hybridization with a PCR-amplifiable synthetic DNA (universal heteroduplex generator, UHG). The UHG contains identifiers consisting of nucleotide substitutions and/or deletions, contiguous with known mutation sites within the target exon. DNA heteroduplexes are resolved by nondenaturing polyacrylamide minigel electrophoresis. Individual mutant genotypes are identified by characteristic banding patterns, in either homozygous or heterozygous states. The method may potentially be applied to rapid genotyping of any mutation or series of mutations within PCR-amplifiable genetic material.
authors.affiliation: University of Bristol Department of Transplantation Sciences, Bristol Homoeopathic Hospital, Cotham, United Kingdom.
authors.forename: N
authors.initials: N
authors.lastname: Wood
authors.forename: L
authors.initials: L
authors.lastname: Tyfield
authors.forename: J
authors.initials: J
authors.lastname: Bidwell
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000138:chemical synthesis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D005069:Evaluation Studies as Topic; D005091:Exons; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D007231:Infant, Newborn; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics; D010651:Phenylalanine Hydroxylase / Q000235:genetics; D010661:Phenylketonurias / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction
chemical_list: D009692:Nucleic Acid Heteroduplexes; D004247:DNA; D010651:Phenylalanine Hydroxylase