title: Mutation screening by a combination of biotin-SSCP and direct sequencing.
abstract: We have developed a mutation detection strategy that combines single strand conformational polymorphism (SSCP) analysis of one strand of a double-stranded amplification product with direct sequencing of the other. Using this strategy, which we find economical of both time and resources, we have identified a G to A transition, which substitutes a serine for glycine residue at position 862 in the major helix of the alpha 1 chain of Type I collagen. We use this mutation, which causes a lethal form of osteogenesis imperfecta, to illustrate the technique.
authors.affiliation: University of Oxford, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, UK.
authors.forename: A S
authors.initials: AS
authors.lastname: Virdi
authors.forename: J A
authors.initials: JA
authors.lastname: Loughlin
authors.forename: C M
authors.initials: CM
authors.lastname: Irven
authors.forename: J
authors.initials: J
authors.lastname: Goodship
authors.forename: B C
authors.initials: BC
authors.lastname: Sykes
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001710:Biotin; D003094:Collagen / Q000235:genetics*; D004247:DNA; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010013:Osteogenesis Imperfecta / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011296:Prenatal Diagnosis; D017422:Sequence Analysis, DNA
chemical_list: D001710:Biotin; D003094:Collagen; D004247:DNA