Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize.
In this study, we developed a novel multiplex polymerase chain reaction (PCR) method for simultaneous 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 an endogenous reference gene, ssIIb, were included in the nonaplex(9plex) PCR system, and its amplified products could be distinguished by agarose gel and capillary electrophoreses based on their different lengths. The optimal condition enabled us to reliably amplify two fragments corresponding to a construct specific sequence and a taxon specific ssIIb in each of the eight events of GM maize and all of nine fragments in a simulated GM mixture containing as little as 0.25% (w/w) each of eight events of GM maize. These results indicate that this multiplex PCR method could be an effective qualitative detection method for screening GM maize.
Onishi
Mari
M
Research and Development Division, Fasmac Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Matsuoka
Takeshi
T
Kodama
Takashi
T
Kashiwaba
Koichi
K
Futo
Satoshi
S
Akiyama
Hiroshi
H
Maitani
Tamio
T
Furui
Satoshi
S
Oguchi
Taichi
T
Hino
Akihiro
A
D001483:Base Sequence; D018744:DNA, Plant; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D003313:Zea mays / Q000235:genetics*
D018744:DNA, Plant