SYBRA®Green qPCR screening methods for the presence of "35S promoter" and "NOS terminator" elements in food and feed products

The Cauliflower Mosaic Virus "35S promotor" (p35S) and the Agrobacterium "Nopaline Synthase" terminator (tNOS) are the most represented generic recombinant elements in commercial genetically modified crops to date. A set of four new SYBR(A (R))Green qPCR methods targeting the "p35S" and "tNOS" core elements have been developed. These qPCR methods generate short amplicons of 147 and 75 bp for the "p35S" element and 172 and 69 bp for the "tNOS" element. Single target plasmids containing these amplicons were constructed and allow determining the nominal melting temperature (T (m) value) of each amplicon. The four methods are specific for their respective targets, and moreover, three of them are highly sensitive (up to 1-2 copies detectable) at a PCR efficiency ranging between 95 and 100%. The latter methods can detect their respective targets at 0.1% (w/w) gDNA levels and are suitable for detecting low levels of genetically modified materials containing the "p35S" and/or "tNOS" elements.