The European Union (EU) imposes strict regulations on the presence of genetically modified (GM) material in food and feed, requiring thorough testing of samples for various GM lines. Although traditional quantitative real-time PCR (qPCR) methods are sensitive and robust, they are not cost-effective for managing large numbers of GM events due to their limited multiplexing capabilities. Conversely, digital PCR (dPCR) is capable of robust quantitative multiplexing in addition to other benefits such as absolute quantification and better tolerance of PCR inhibitors. In this context, we present a protocol for multiplex quantification of 19 GM soybean lines using dPCR as an improvement over the currently used simplex qPCR approach. This method enables simple and robust quantification of common GM soybean lines with a relatively low number of reactions.

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Multiplex Quantification of 19 GM Soybean Lines Using Digital PCR

  • Amadej Jelenčič,
  • Dejan Štebih,
  • Tina Demšar,
  • David Dobnik

摘要

The European Union (EU) imposes strict regulations on the presence of genetically modified (GM) material in food and feed, requiring thorough testing of samples for various GM lines. Although traditional quantitative real-time PCR (qPCR) methods are sensitive and robust, they are not cost-effective for managing large numbers of GM events due to their limited multiplexing capabilities. Conversely, digital PCR (dPCR) is capable of robust quantitative multiplexing in addition to other benefits such as absolute quantification and better tolerance of PCR inhibitors. In this context, we present a protocol for multiplex quantification of 19 GM soybean lines using dPCR as an improvement over the currently used simplex qPCR approach. This method enables simple and robust quantification of common GM soybean lines with a relatively low number of reactions.