Patent ID: 9096896
Filing Date: 2015-08-04
Classification: C12Q

Abstract:
1. A method of quantifying event Bt11 DNA in a biological sample comprising corn nucleic acids, the method comprising: (a) contacting said biological sample with a first pair of primers, comprising a first primer consisting of SEQ ID NO: 1 and a second primer consisting of SEQ ID NO: 2, and a fluorescent dye labeled probe comprising SEQ ID NO: 3, wherein said first pair of primers, when used in a nucleic-acid amplification reaction with genomic DNA from event Bt11 corn, produce a first amplicon comprising SEQ ID NO: 4, and wherein said first amplicon is diagnostic for event Bt11; (b) contacting said biological sample with a second pair of primers, comprising a first primer consisting of SEQ ID NO: 5 and a second primer consisting of SEQ ID NO: 6, and a second fluorescent dye labeled probe comprising SEQ ID NO: 7, wherein said second pair of primers, when used in a nucleic-acid amplification reaction with corn genomic DNA, produce a second amplicon comprising SEQ ID NO: 8, and wherein said second amplicon is indicative of the presence of a maize adh1 gene; (c) providing a nucleic acid amplification reaction condition and a PCR instrument capable of performing a quantitative real-time PCR; (d) performing the quantitative real-time PCR using the primers and probes of (a) and (b), thereby producing said first amplicon and said second amplicon; (e) detecting simultaneously said first amplicon and said second amplicon as they are produced by said PCR instrument; and (f) calculating a relative amount of said first amplicon compared to said second amplicon, whereby the amount of said first amplicon is indicative of the quantity of Bt11 DNA in said biological sample.