Simultaneous visual detection of single-nucleotide variations in tuna DNA using DNA/RNA chimeric probes and ribonuclease A

研究成果: ジャーナルへの寄稿記事

6 引用 (Scopus)

抄録

The need for detection of single-nucleotide polymorphisms (SNPs) is rapidly increasing for molecular diagnostics, species authentication, and food traceability. In many detection technologies, fluorescence probes have the advantage of simultaneous detection of multiple analytes using multiple color fluorescence dyes. In addition, an adequate concentration of fluorescence can be observed by the naked eye. We conducted a visual ribonuclease protection assay using multicolor fluorescence probes for the simultaneous detection of multiple tuna species. The assay includes amplification of a target RNA sequence by in vitro transcription, hybridization with DNA/RNA chimeric fluorescence probes, and cleavage of mismatched RNA bases by ribonuclease A. Fragmented dye-labeled oligonucleotides were easily removed by centrifugal gel filtration. Using three fluorescent probes, fluorescence signals related to the target SNP were simply found by the visual observation of eluates. Moreover, the dual fluorescence system was used for obtaining the mixing ratio of two tuna species. This technique appeared to be convenient for detection of interest species in food products.

元の言語英語
ページ(範囲)6-11
ページ数6
ジャーナルAnalytical Biochemistry
389
発行部数1
DOI
出版物ステータス出版済み - 6 1 2009

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Tuna
RNA Probes
Pancreatic Ribonuclease
Nucleotides
Fluorescence
RNA
DNA
Polymorphism
Single Nucleotide Polymorphism
Assays
Coloring Agents
RNA Cleavage
Food
Molecular Pathology
Transcription
Ribonucleases
Fluorescent Dyes
Oligonucleotides
Authentication
Gel Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

これを引用

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abstract = "The need for detection of single-nucleotide polymorphisms (SNPs) is rapidly increasing for molecular diagnostics, species authentication, and food traceability. In many detection technologies, fluorescence probes have the advantage of simultaneous detection of multiple analytes using multiple color fluorescence dyes. In addition, an adequate concentration of fluorescence can be observed by the naked eye. We conducted a visual ribonuclease protection assay using multicolor fluorescence probes for the simultaneous detection of multiple tuna species. The assay includes amplification of a target RNA sequence by in vitro transcription, hybridization with DNA/RNA chimeric fluorescence probes, and cleavage of mismatched RNA bases by ribonuclease A. Fragmented dye-labeled oligonucleotides were easily removed by centrifugal gel filtration. Using three fluorescent probes, fluorescence signals related to the target SNP were simply found by the visual observation of eluates. Moreover, the dual fluorescence system was used for obtaining the mixing ratio of two tuna species. This technique appeared to be convenient for detection of interest species in food products.",
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