mericon GMO Screen 35S-pat Kit
For real-time PCR detection of GMOs with the 35S-pat construct in food or animal feed
The mericon GMO Screen 35S-pat Kit is designed for the target-specific detection of the 35S-pat construct DNA in food, animal feed, and pharmaceutical products. The assay uses a real-time PCR-based protocol and forms part of the comprehensive QIAGEN food testing portfolio, which also features dedicated sample preparation kits and assays for pathogen detection and ingredient authentication. The mericon GMO Screen 35S-pat Kit performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.
The mericon GMO Screen 35S-pat Kit is intended for molecular biology applications in food, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.
The mericon GMO Screen 35S-pat Kit is for the detection of the 35S construct in food and animal feed. This construct links the pat gene from Streptomyces viridochromogenes to the 35S promoter of cauliflower mosaic virus. The promoter triggers the expression of phosphinotricin-acetyltransferase, which confers resistance to the herbicide phosphinotricin. The assay can detect as few as 10 copies of the target DNA in a reaction and it exhibits high specificity for the 35S-pat construct. In an experiment using 50 ng of tested DNA, no cross-reactivity was observed with other genetic constructs or with plant or animal species not containing this genetic construct (see the table "Results of mericon GMO Screen 35S-pat Kit cross-reactivity study").
Results of mericon GMO Screen 35S-pat Kit cross-reactivity study
The mericon GMO Screen 35S-pat Kit is designed for the qualitative detection of genetically modified organisms containing the pat gene, driven by the 35S promoter. The expression of the pat gene mediates tolerance to the herbicide phosphinotricin in crops.
Detection of GMOs using real-time PCR is based on the amplification of a specific region of the relevant GMO genome. The amplified product is detected using target-specific fluorescent probes that bind to it. As the PCR product accumulates, there is 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.
mericon PCR Assays include an internal control to control potential inhibition. This aids in the straightforward interpretation of the results (see the table "Summary of possible PCR outcomes").
The PCR primer set for each assay is highly specific and targets either a genetic construct that is commonly used in GMOs or a DNA sequence that is specific for a particular GMO. These targets were verified bioinformatically and experimentally.
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 wide 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 GMO Screen 35S-pat Kit is for real-time PCR-based detection of GMOs with the 35S-pat construct in food, animal feed, and pharmaceutical products.
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