Fluorogenic ribonuclease protection (FRIP) analysis of single nucleotide polymorphisms (SNPs) in Japanese rice (Oryza sativa L.) DNA for cultivar discrimination

Momoko Kitaoka, Takuya Wada, Takeshi Nishio, Masahiro Goto

    研究成果: ジャーナルへの寄稿学術誌査読

    2 被引用数 (Scopus)

    抄録

    A rapid and easy method to discriminate plant cultivars is indispensable to confirm food labeling. We established a fluorogenic ribonuclease protection (FRIP) assay to discriminate Japanese rice (Oryza sativa L.) cultivars based on single nucleotide polymorphisms (SNPs). The FRIP assay uses a hybridization technique between fluorescent probes and the target sequence prepared by run-off transcription, but the requirement of two PCR thermocycles is the problem when preparing template DNA for run-off transcription from rice genomic DNA. In this study, we designed new PCR primers with asymmetric melting temperatures. These primers amplified the target SNP marker containing a T7 RNA polymerase promoter sequence upstream of the target sequence in a single PCR. Moreover, 100 cultivars were discriminated with the patterns of 15 SNPs. The assay can be used as a rapid method of analysis to discriminate Japanese rice cultivars.

    本文言語英語
    ページ(範囲)2189-2193
    ページ数5
    ジャーナルBioscience, Biotechnology and Biochemistry
    74
    11
    DOI
    出版ステータス出版済み - 2010

    !!!All Science Journal Classification (ASJC) codes

    • バイオテクノロジー
    • 分析化学
    • 生化学
    • 応用微生物学とバイオテクノロジー
    • 分子生物学
    • 有機化学

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