Fluorogenic Ribonuclease Protection (FRIP) Analysis of Single Nucleotide Polymorphisms (SNPs) in Japanese Rice (Oryza sativa L.) DNA for Cultivar Discrimination
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概要
- 論文の詳細を見る
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.
- 社団法人 日本農芸化学会の論文
- 2010-11-23
著者
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GOTO Masahiro
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University
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Goto Masahiro
Department Of Applied Chemistry Graduate School Of Engineering Kyushu University
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Wada Takuya
Department Of Crop Science Fukuoka Agricultural Research Center
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Nishio Takeshi
Graduate School of Agric. Sci., Tohoku Univ.
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Nishio Takeshi
Graduate School Of Agricultural Science Tohoku University
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Nishio Takeshi
Graduate School Of Agric. Sci. Tohoku Univ.
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KITAOKA Momoko
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University
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Kitaoka Momoko
Department Of Applied Chemistry Graduate School Of Engineering Kyushu University
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Goto Masahiro
Department Of Applied Biological Chemistry The University Of Tokyo
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