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mericon GMO Screen 35S-pat Kit

For real-time PCR detection of GMOs with the 35S-pat construct in food or animal feed

  • Highly sensitive and specific detection of GMOs with the 35S-pat construct
  • Streamlined detection protocol
  • Dedicated mericon DNA isolation kit available
  • PCR run time of 82 minutes on the QIAGEN Rotor-Gene Q
  • Full license for PCR without additional costs
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.
Cat No./ID: 291023
mericon GMO Screen 35S-pat Kit (24)
$254.00
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For 24 reactions: PCR Assay Screen 35S-pat, Internal Control, Positive Control, Multiplex PCR Master Mix, QuantiTect Nucleic Acid Dilution Buffer, RNase-free water, 50x ROX Dye Solution
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.

Performance

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

 Species, GMO, or
 genetic construct
 Result  Species, GMO or 
 genetic construct
 Result
 Bt11 corn  +  Bt176 corn  –
 TC1507 corn  +  EH92-527 potato  –
 59122 corn  +  H7-1 sugar beet  –
 98140 corn  –  356043 soy  –
 3272 corn  –  Soy  –
 NK603 corn  –  Wheat  –
 MIR604corn  –  Rye  –
 MON863 corn  –  Barley  –
 MON810 corn  –  Pig  –
 98140 corn  –  Sheep  –
 Roundup Ready soy  –  Goat  –
 305423 soy  –  Cattle  –

Principle

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. 

Summary of possible PCR outcomes
Internal control amplifiedTested DNA amplified Meaning
            +            +  Sample is positive for the target DNA
            +            –  Sample is negative for the target DNA
            –            –  PCR failed due to inhibition
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 35S-pat Kit is for real-time PCR-based detection of GMOs with the 35S-pat construct in food, animal feed, and pharmaceutical products.
Features
Specifications
Quantitative/qualitative Qualitative
Recommended sample prep DNeasy mericon Food Kit or DNeasy Plant Kits
RUO/CE/ASR RUO
Sample type Food
Thermal cycler Rotor-Gene Q and other cyclers
What detected GMO DNA
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