Detection of single-base mutations by fluorogenic ribonuclease protection assay

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

研究成果: ジャーナルへの寄稿記事

16 引用 (Scopus)

抄録

The ribonuclease protection assay is a generally applicable technique for the detection of known mutations. We have developed a simple and rapid method for mutation detection based on the ribonuclease protection assay using fluorescently labeled oligodeoxyribonucleotide probes. The fluorogenic ribonuclease protection (FRAP) assay uses two differently labeled oligodeoxyribonucleotides, a donor probe and an acceptor probe, to obtain a fluorescence resonance energy transfer (FRET) signal. We have utilized the FRAP assay for the detection of a single-base mutation in the YMDD motif of the hepatic B virus DNA polymerase gene. The occurrence of mismatch-selective RNA cleavage was successfully discriminated by measuring the FRET signal between the donor and acceptor probes. Moreover, mutation sensing was successfully visualized by a UV transillumination. This simple and rapid mutation sensing method should facilitate a high-throughput mutation analysis.

元の言語英語
ページ(範囲)7047-7053
ページ数7
ジャーナルAnalytical Chemistry
77
発行部数21
DOI
出版物ステータス出版済み - 11 1 2005

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Ribonucleases
Assays
Transillumination
Oligonucleotide Probes
Oligodeoxyribonucleotides
DNA-Directed DNA Polymerase
Viruses
Genes
Throughput
RNA
Fluorescence Resonance Energy Transfer

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

これを引用

Detection of single-base mutations by fluorogenic ribonuclease protection assay. / Ichinose, Hirofumi; Kitaoka, Momoko; Okamura, Nobuko; Maruyama, Tatsuo; Kamiya, Noriho; Goto, Masahiro.

:: Analytical Chemistry, 巻 77, 番号 21, 01.11.2005, p. 7047-7053.

研究成果: ジャーナルへの寄稿記事

Ichinose, Hirofumi ; Kitaoka, Momoko ; Okamura, Nobuko ; Maruyama, Tatsuo ; Kamiya, Noriho ; Goto, Masahiro. / Detection of single-base mutations by fluorogenic ribonuclease protection assay. :: Analytical Chemistry. 2005 ; 巻 77, 番号 21. pp. 7047-7053.
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AU - Goto, Masahiro

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