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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Related Experiment Video

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A PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica
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Simple, fast, reliable: multiplex digital PCR quantification of 19 genetically modified soybean events.

Amadej Jelenčič1,2, Dejan Štebih1, Tina Demšar1

  • 1Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.

GM Crops & Food
|February 28, 2026
PubMed
Summary

A new multiplex digital PCR (dPCR) method enables high-throughput quantification of 19 genetically modified (GM) soybean events. This advanced technique supports regulatory compliance for GM crops with a simplified workflow.

Keywords:
Genetically modified (GM) cropsGlycine max (L.) merrdigital PCR (dPCR)multi-targetmultiplexquantificationsoybeantransgenic

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Area of Science:

  • Agricultural biotechnology
  • Molecular biology
  • Analytical chemistry

Background:

  • Modern agriculture increasingly utilizes plant genetic engineering, leading to a rise in genetically modified (GM) crop varieties.
  • Regulatory bodies require high-throughput, multi-target analytical methods for detecting and quantifying GM content in crops.
  • Existing methods may lack the comprehensiveness or efficiency needed for routine monitoring.

Purpose of the Study:

  • To develop and validate a multiplex digital PCR (dPCR) method for the simultaneous detection and quantification of multiple GM soybean events.
  • To establish a high-throughput analytical approach suitable for regulatory compliance and routine GM crop monitoring.
  • To create a comprehensive quantification strategy for GM soybeans that simplifies workflow and promotes widespread adoption.

Main Methods:

  • Development of four 5-plex digital PCR (dPCR) assays for simultaneous detection of 19 GM soybean events and the lectin (Le1) endogenous gene.
  • Utilization of a nanowell plate-based, all-in-one dPCR system with multiple fluorescence detection channels.
  • Validation of assays against minimum performance requirements including specificity, trueness, precision, sensitivity, and dynamic range.

Main Results:

  • The developed multiplex dPCR method successfully quantified 19 GM soybean events and the Le1 endogene.
  • The four 5-plex assays met all minimum performance requirements for routine GM crop analysis.
  • This represents the most comprehensive multi-target GM soybean quantification method to date, requiring no prior screening.

Conclusions:

  • The presented multiplex dPCR method offers a robust, efficient, and comprehensive solution for GM soybean quantification.
  • The simplified workflow and validated performance make the method suitable for widespread adoption in regulatory settings.
  • This study establishes a precedent for developing rapid, multiplex dPCR assays for evolving GM crop monitoring demands.