Mutation detection in DNA oligonucleotides based on a guanine quenching method coupled with enzymatic digestion of single-stranded DNA

Tatsuo Maruyama, Toshimitsu Shinohara, Hirofumi Ichinose, Momoko Kitaoka, Nobuko Okamura, Noriho Kamiya, Masahiro Goto

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Fluorescence quenching by guanine allows DNA hybridization to be monitored and any point mutations in oligonucleotides to be detected. However, fluorescence quenching is often affected by untargeted guanine located in a protruding end (single-strand DNA) of the probe-target DNA duplex resulting in an unsatisfactory sensitivity. In the present study, we used enzymatic digestion of the protruding end of a probe-target DNA duplex to avoid interference by untargeted guanine on fluorescence quenching for detection of a nucleobase mutation. Enzymatic digestion of the protruding end of the DNA duplex fully prevented interference by untargeted guanine, and produced a marked difference in the quenching ratios (36% for wild-type, and 0% for mutant).

Original languageEnglish
Pages (from-to)1349-1354
Number of pages6
JournalBiotechnology letters
Volume27
Issue number18
DOIs
Publication statusPublished - Sep 1 2005

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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