mericon Quant MON 810 Kit

The mericon Quant MON 810 Kit is designed for the specific detection and quantification of MON 810 corn DNA in food and animal feed. Both the GMO-specific MON 810 Assay and the Reference System Assay (corn assay) can detect as few as 10 copies of target DNA in a reaction.

The PCR Assay MON 810 exhibits high specificity for MON 810 corn. No cross-reactivity was observed with other plant and animal species, GMOs, or genetic constructs not found in MON 810 corn using 50 ng of tested DNA (see the table "Results of mericon Quant MON 810 Corn Kit cross-reactivity study").

Results of mericon Quant MON 810 Corn Kit cross-reactivity study

The Reference System Assay (corn assay) exhibits high specificity for corn and MON 810 corn. No cross-reactivity was observed with other plant or animal species or GMOs in an experiment using 50 ng of tested DNA (see the table "Results of Reference System Assay cross-reactivity study").

Results of Reference System Assay cross-reactivity study

The mericon Quant MON 810 Corn Kit was designed for the quantitavite detection of the genetically modified organism (GMO) MON 810 corn (one of the most frequently used genetically modified corn lines worldwide) in food and animal feed samples. The corn line MON 810 was developed through a genetic modification resulting in resistance against the European corn borer (Ostrinia nubilalis) by producing Bt‑Toxins of Bacillus thuringiensis.

Detecting DNA from a genetically modified organism (GMO) via PCR is based on amplifying a genetic construct used in that GMO. In real-time PCR, the amplified product is detected via target-specific fluorescent probes that bind to the amplified product. Accumulation of PCR product results in an increased fluorescent signal from the bound probes. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating PCR product without having to re-open the reaction tubes afterward.

The probes of mericon PCR Assays are sequence-specific oligonucleotides with a fluorophore at the 5' end of the probe, and a quencher moiety at the 3' end. If the target DNA sequence is present, the probe is cleaved by the 5'→3' exonuclease activity of HotStarTaq Plus DNA Polymerase during the extension phase of PCR. This separates the fluorophore and the quencher moiety, resulting in a detectable fluorescence that is proportional to the amount of accumulated PCR product.

The target GMO is quantified based on the alignment of the PCR results from a GMO-specific PCR system and a Reference PCR system. The GMO assay targets either a genetic construct commonly used in a desired GMO or a DNA sequence that is specific for a particular GMO (e.g., Roundup Ready Soy). The Reference assay targets the plant species of the GMO (e.g., soy). Standard curves for both systems form the basis of the quantification experiment. They are generated with a defined dilution of the Standard DNA, which contains a defined ratio of GMO and Reference DNA.

Each mericon Quant Assay contains an optimized mixture of PCR primer sets for a GMO-specific and a species-specific (Reference System) target sequence, each with an internal control (IC), plus probes labeled with distinct fluorescent dyes (in both systems, the test sample is detected with FAM reporter (495/520 nm), and the internal control is detected with MAX NHS Ester reporter (MAX; 524/557 nm)). In addition, each kit includes a Quantification Control DNA, a Standard DNA, and all of the reagents necessary to perform the analysis.

DNA is extracted from a sample of food or animal feed using a dedicated sample preparation kit. QIAGEN's DNeasy mericon Food Kit is suitable for a whole range of food matrices, including those with high inhibitor contents.

The isolated, pure DNA is assayed using a streamlined real-time PCR protocol that returns results in 82 minutes on QIAGEN's Rotor-Gene Q. The protocol has also been validated for use on other thermal block cyclers.

The results are displayed as easy-to-interpret PCR curves.

The mericon Quant MON 810 Corn Kit is for real-time PCR-based detection and quantification of MON 810 corn in food, animal feed, and pharmaceutical products.