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 re-sources. SYBR^® Green I-based duplex and triplex re-al-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 sim-ultaneously identify multiple events. Duplex real-time PCR assay based on TaqMan^® chemistry targeting Bt176 event-specific sequence and Adh1 was also op-timized for quantification purpose. Limit of detection of developed assays was up to 0.05% and limit of quantification of the reported TaqMan^® based real-time PCR was up to 0.5%.