Multiplex real-time PCR assays for simultaneous detection of maize MON810 and GA21 in food samples

This work describes a quantitative multiplex real-time PCR method optimized for the detection 12 of maize MON810 and GA21. The use of specific primers and of labeled probes by real-time PCR 13 allowed for the simultaneous detection and confirmation of amplicon identity and increased the 14 reliability of the technique and the number of PCR applications to food analysis. 15 Two different endogenous genes, Zein and Adh1, were evaluated for quantitative use as accountable for continuous development of maize traits. The quantification is based on a calibration standard curve obtained with the DNA extracted from Certified Reference Materials (CRMs). The limit of detection (LOD) and limit of quantification (LOQ) of the triplex assays developed was set at 3 and 36 copy numbers respectively.