Multiplex real-time PCR-based detection and quantification of genetically modified maize events employing SYBR® Green I and TaqMan® chemistries

Multiplexing in real-time PCR offers advantages over singleplex real-time PCR by saving time and resources. SYBR (R) Green I-based duplex and triplex real-time PCR assays targeting event-specific sequences of three genetically modified (GM) maize events, namely Bt11, Bt176 and MON89034, and taxon-specific endogenous Adh1 gene were developed to simultaneously identify multiple events. Duplex real-time PCR assay based on TaqMan (R) chemistry targeting Bt176 event-specific sequence and Adh1 was also optimized for quantification purpose. Limit of detection of developed assays was up to 0.05% and limit of quantification of the reported TaqMan (R) based real-time PCR was up to 0.5%.