mericon GMO Screen bar Kit
For real-time PCR detection of the bar gene in food or animal feed
The mericon GMO Screen bar Kit is designed for the target-specific detection of the bar gene 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 bar Kit performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.
The mericon GMO Screen bar 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.
Performance
The mericon Screen bar Kit is designed for the detection of the bar gene in food and animal feed. The mericon Screen bar Kit can detect as little as 10 copies of target DNA in a reaction and exhibits high specificity for the bar gene. No cross-reactivity was observed with other genetic constructs or with plant or animal species not containing this genetic construct using 50 ng of tested DNA (see the table "Results of mericon GMO Screen bar Kit cross-reactivity study"). Results of mericon GMO Screen bar Kit cross-reactivity study
Principle
The mericon GMO Screen bar Kit is designed for the qualitative detection of genetically modified organisms (GMOs) containing the bar gene. The gene, which originates from Streptomyces hygroscopicus, transcripts phosphinotricin N-Acetyl-transferase and confers resistance to the active compound phosphinotricin of herbicides. 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.
Procedure
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.
Applications
The mericon GMO Screen bar Kit is for real-time PCR-based detection of GMOs with the bar gene in food, animal feed, and pharmaceutical products.
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