Premium
Mass Spectrometric Gene Diagnosis of One‐base Substitution from Polymerase Chain Reaction Amplified Human DNA
Author(s) -
Tsuneyoshi Toshihiro,
Ishikawa Keiichiro,
Koga Yoshinori,
Naito Yasuhiro,
Baba Shozo,
Terunuma Hideya,
Arakawa Ryuichi,
Prockop Darwin J.
Publication year - 1997
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19970422)11:7<719::aid-rcm862>3.0.co;2-j
Subject(s) - chemistry , ecori , adduct , dna , chloroform , mass spectrometry , polymerase chain reaction , electrospray ionization , restriction enzyme , microbiology and biotechnology , chromatography , gene , biochemistry , organic chemistry , biology
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 Eco RI 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. © 1997 John Wiley & Sons, Ltd.