Patent ID: 8685677

Claim:
A method for determining event zygosity of a soybean plant comprising an AAD-12 soybean event pDAB4468-0416, said event comprising a transgene construct comprising an AAD-12 gene, said transgene construct being flanked by a 5â€² flanking soybean genomic DNA and a 3â€² flanking soybean genomic DNA, said method comprising: obtaining a DNA sample of genomic DNA from said soybean plant; producing a contacted sample by contacting said DNA sample with a. a first event primer and a second event primer, wherein said first event primer specifically binds said transgene construct, said second event primer specifically binds said 5â€² soybean genomic flanking DNA or said 3â€² soybean genomic flanking DNA, and wherein said first event primer and said second event primer produce an event amplicon when subjected to TAQMAN PCR conditions; b. a reference forward primer and a reference reverse primer that produce a reference amplicon from an endogenous soybean reference gene when subjected to TAQMAN PCR conditions; c. a florescent event probe that hybridizes with said event amplicon; d. a florescent reference probe that hybridizes with said reference amplicon; subjecting said contacted sample to fluorescence-based endpoint TAQMAN PCR conditions; quantitating said florescent event probe that hybridized to said event amplicon; quantitating said florescent reference probe that hybridized to said reference amplicon; comparing amounts of hybridized florescent event probe to hybridized florescent reference probe; and determining zygosity of pDAB4468-0416 by comparing florescence ratios of hybridized fluorescent event probe and hybridized fluorescent reference probe.