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

研究成果: Contribution to journalArticle査読

20 被引用数 (Scopus)

抄録

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).

本文言語英語
ページ(範囲)1349-1354
ページ数6
ジャーナルBiotechnology letters
27
18
DOI
出版ステータス出版済み - 9 1 2005

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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