Mass spectrometric gene diagnosis of one-base substitution from polymerase chain reaction amplified human DNA

Toshihiro Tsuneyoshi, Keiichiro Ishikawa, Yoshinori Koga, Yasuhiro Naito, Shozo Baba, Hideya Terunuma, Ryuichi Arakawa, Darwin J. Prockop

Research output: Contribution to journalArticle

28 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)719-722
Number of pages4
JournalRapid Communications in Mass Spectrometry
Volume11
Issue number7
DOIs
Publication statusPublished - 1997

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

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