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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

Updated: Oct 17, 2025

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres
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Simple, Precise, and Less Biased GMO Quantification by Multiplexed Genetic Element-Specific Digital PCR.

Satoshi Noma1, Yosuke Kikuchi1, Megumi Satou2

  • 1Research Center for Basic Science, Nisshin Seifun Group Inc, 5-3-1 Tsurugaoka, Fujimino, Saitama 356-8511, Japan.

Journal of AOAC International
|October 9, 2021
PubMed
Summary

A new digital PCR method simplifies genetically modified organism (GMO) quantification in maize. This approach offers a more precise and less biased way to ensure accurate GMO food labeling as GM events increase globally.

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Area of Science:

  • Agricultural Biotechnology
  • Molecular Biology
  • Food Science

Background:

  • Global food labeling regulations require accurate quantification of genetically modified organisms (GMOs).
  • Distinguishing between GMO and non-GMO products necessitates reliable GMO content measurement methods.
  • The increasing prevalence of GMOs worldwide drives the need for advanced detection strategies.

Purpose of the Study:

  • To develop a cost-effective, simple, and accurate analytical strategy for GMO detection.
  • To establish a digital PCR-based method for quantifying GMO content in maize grains.
  • To address the growing need for reliable GMO quantification methods due to the increasing number of GM events.

Main Methods:

  • Development of a multiplex digital PCR (dPCR) assay targeting common GMO genetic elements (35S promoter and nopaline synthase terminator).
  • Application of dPCR for quantifying GMO content in maize grain samples.
  • Comparison of the dPCR method with conventional quantitative real-time PCR (qPCR) techniques.

Main Results:

  • The developed multiplex dPCR method is simpler and more precise than conventional qPCR methods.
  • The method accurately quantifies GMO DNA copy numbers, avoiding bias from tandemly integrated genetic elements.
  • Demonstrated ability to quantify copy number of GMO DNA without double counting tandemly placed elements.

Conclusions:

  • This study presents the first multiplexed genetic element-specific digital PCR method for quantifying genetically modified maize.
  • The developed method effectively reduces analytical bias, particularly with tandemly integrated genetic elements.
  • Multiplexed digital PCR offers a simplified approach for GMO quantification, crucial for evolving GM event landscapes.