title: Mass spectrometric gene diagnosis of one-base substitution from polymerase chain reaction amplified human DNA.
abstract: One-base substitution has been detected on the polymerase chain reaction (PCR) products amplified from human mutated DNA for the first time by using mass spectrometry. PCR fragments of 52 base pairs were produced on a collagen gene of an osteogenesis imperfecta patient's heterozygous DNAs. The products were digested with EcoRI restriction enzyme to liberate 3'-end adducts and purified by phenol + chloroform extraction, ammonium acetate addition and ethanol precipitation to remove sodium ions from the phosphoric acid backbone of the DNAs. Purified products were examined using an electrospray ionization mass spectrometer. Mass spectra showed four groups of fragment peaks with the expected molecular masses, which originate from the sense and antisense strands of the heterozygous DNAs.
authors.affiliation: Department of Materials Science, Shizuoka Institute of Science and Technology, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Tsuneyoshi
authors.forename: K
authors.initials: K
authors.lastname: Ishikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Koga
authors.forename: Y
authors.initials: Y
authors.lastname: Naito
authors.forename: S
authors.initials: S
authors.lastname: Baba
authors.forename: H
authors.initials: H
authors.lastname: Terunuma
authors.forename: R
authors.initials: R
authors.lastname: Arakawa
authors.forename: D J
authors.initials: DJ
authors.lastname: Prockop
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D016678:Genome; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012964:Sodium / Q000737:chemistry
chemical_list: D004247:DNA; D012964:Sodium