Transgenic Corn Event MON95275 and Methods for Detection and Uses Thereof

A method of determining the zygosity of a corn plant or corn seed comprising corn event MON95275 comprising:

• a) contacting a sample comprising corn DNA with a first primer pair that is capable of producing an amplicon diagnostic for the allele corresponding to corn event MON95275 DNA;
• b) contacting said sample with a second primer pair that is capable of producing, using a thermal amplification reaction, an amplicon of an internal standard corn genomic DNA known to be single-copy and homozygous in the corn plant;
• c) contacting said sample with a probe set which contains at least a first probe that specifically hybridizes to (or with) the allele DNA of corn event MON95275, and a second probe that specifically hybridizes to the internal standard corn genomic DNA known to be single-copy and homozygous in the corn plant;
• d) performing a DNA amplification reaction using real-time PCR and determining the cycle thresholds (Ct values) of the amplicon corresponding to the allele DNA of corn event MON95275 and the single-copy, homozygous internal standard;
• e) calculating the difference (ΔCt) between the Ct value of the single-copy, homozygous internal standard amplicon and the Ct value of the amplicon corresponding to the allele DNA of corn event MON95275;
• and
• f) determining zygosity, wherein a ΔCt of about zero (0) indicates homozygosity of the inserted T-DNA of event MON95275 and a ΔCt of about one (1) indicates heterozygosity of the inserted T-DNA of corn event MON95275.

A method of determining the zygosity of a corn plant or corn seed comprising corn event MON95275 comprising:

• a) contacting a sample comprising corn DNA with a set of primer pairs comprising at least two different primer pairs capable of producing a first amplicon diagnostic for corn event MON95275 and a second amplicon diagnostic for native corn genomic DNA devoid of corn event MON95275 DNA;
• b) performing a nucleic acid amplification reaction with the sample and the set of primer pairs;
• and
• c) detecting in the nucleic acid amplification reaction the first amplicon diagnostic for the presence of corn event MON95275 DNA, or the second amplicon diagnostic for native corn genomic DNA devoid of corn event MON95275, wherein the presence of only the first amplicon is diagnostic of a corn plant or corn seed homozygous for corn event MON95275 DNA, and the presence of both the first amplicon and the second amplicon is diagnostic of a corn plant or corn seed heterozygous for corn event MON95275 DNA;
• or
• d) contacting a sample comprising corn DNA with a probe set which contains at least a first probe that specifically hybridizes to corn event MON95275 DNA and at least a second probe that specifically hybridizes to corn genomic DNA that was disrupted by insertion of the heterologous DNA of corn event MON95275 and does not hybridize to corn event MON95275 DNA;
• and
• e) hybridizing the probe set with the sample under stringent hybridization conditions,
• wherein detecting hybridization of only the first probe under the hybridization conditions is diagnostic for a homozygous allele of corn event MON95275 DNA, and wherein detecting hybridization of both the first probe and the second probe under the hybridization conditions is diagnostic for a corn plant or seed heterozygous for corn event MON95275 DNA.