PCR-Based Detection and Quantification of a Transgenic Glyphosate-Tolerant Canola Using a Novel Reference Gene System

Development and commercialization of a transgenic crop depends upon the availability of a reliable quantitative assay sensitive enough to detect low level presence of transgenic material in mixed seed or in a commercial product. An appropriate species-specific endogenous control also is necessary for accurate quantification. A novel mode of transgenic tolerance to glyphosate, whereupon an engineered acetylase detoxifies glyphosate, was recently developed and incorporated into transgenic canola (Brassica napus). Here, we describe two highly specific and sensitive PCR-based assays for the transgenic event, DP-73496-4. We also developed and evaluated an appropriate endogenous control to be used in conjunction with the event-specific assays. This endogenous control is specific to the A genome of cultivated canola-quality oilseed rape based on our analysis of the diversity of FatA gene sequences.