Event detection AAD-1 DAS 40278-9

A method for determining the zygosity of a plant comprising corn event AAD-1 DAS-40278-9 maize, said event comprising a transgene construct comprising an AAD-1 gene, said transgenic construct flanked by genomic DNA being flanking maize 5 'and flanking maize genomic DNA 3', said method comprising: obtaining a DNA sample of genomic DNA from said corn plant; producing a sample contact by contacting said DNA sample with a. a first primer and a second primer event of the event, wherein the event said first primer binds specifically to said transgene construct as defined by the 1874-6689 residues of SEQ ID NO: 29, said second primer event specifically binds said genomic DNA flanking maize 5 'as defined by 1-1873 residues in SEQ ID NO: 29 or said genomic DNA flanking corn 3' as defined by residues 6690-8557 of SED ID NO: 29, and wherein said first primer and said second primer event event produce an amplicon event when subjected to a PCR reaction b. a forward reference and a reverse primer of reference that produce an amplicon reference from an endogenous reference gene of maize when subjected to a PCR reaction, wherein said reference gene: i. It is an endogenous invertase gene Zea mays; or ii. comprises a sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6 and SEQ ID NO: 7 or a sequence having at least 90% sequence identity with any of the sequences mentioned above, c. a fluorescent probe that hybridizes with the event said amplicon event, d. a fluorescent reference probe which hybridizes to said amplicon reference; subjecting said sample to a contact probe assay based on fluorescence hydrolysis which liberates the fluorescent moiety of the probe of the fluorescence quencher moiety of the probe; quantizing said fluorescent probe has hybridized to event event said amplicon; quantizing said fluorescent probe reference amplicon hybridized to said reference; comparing the amount of hybridized fluorescent probe event with the fluorescent probe hybridized reference; and determine the zygosity of DAS-40278-9 by comparing rates of fluorescence of the hybridized fluorescent probe event and hybridized fluorescent probe reference.