Development of a Multiplex Polymerase Chain Reaction Method for Simultaneous Detection of Eight Events of Genetically Modified Maize

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• 作者：Mari Onishi ; Takeshi Matsuoka ; Takashi Kodama ; Koichi Kashiwaba ; Satoshi Futo ; Hiroshi Akiyama ; Tamio Maitani ; Satoshi Furui ; Taichi Oguchi ; and Akihiro Hino
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2005
• 出版时间：December 14, 2005
• 年：2005
• 卷：53
• 期：25
• 页码：9713 - 9721
• 全文大小：583K
• 年卷期：v.53,no.25(December 14, 2005)
• ISSN：1520-5118

文摘

In this study, we developed a novel multiplex polymerase chain reaction (PCR) method forsimultaneous detection of up to eight events of genetically modified (GM) maize within a single reaction.The eight detection primer pairs designed to be construct specific for eight respective GM events(i.e., Bt11, Event176, GA21, MON810, MON863, NK603, T25, and TC1507) and a primer pair for anendogenous reference gene, ssIIb, were included in the nonaplex(9plex) PCR system, and its amplifiedproducts could be distinguished by agarose gel and capillary electrophoreses based on their differentlengths. The optimal condition enabled us to reliably amplify two fragments corresponding to aconstruct specific sequence and a taxon specific ssIIb in each of the eight events of GM maize andall of nine fragments in a simulated GM mixture containing as little as 0.25% (w/w) each of eightevents of GM maize. These results indicate that this multiplex PCR method could be an effectivequalitative detection method for screening GM maize.Keywords: Multiplex PCR; genetically modified (GM); maize; Bt11; Event176; GA21; MON810; MON863;NK603; T25; TC1507; ssIIb