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GMO quantification: valuable experience and insights for the future

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Abstract

Cultivation and marketing of genetically modified organisms (GMOs) have been unevenly adopted worldwide. To facilitate international trade and to provide information to consumers, labelling requirements have been set up in many countries. Quantitative real-time polymerase chain reaction (qPCR) is currently the method of choice for detection, identification and quantification of GMOs. This has been critically assessed and the requirements for the method performance have been set. Nevertheless, there are challenges that should still be highlighted, such as measuring the quantity and quality of DNA, and determining the qPCR efficiency, possible sequence mismatches, characteristics of taxon-specific genes and appropriate units of measurement, as these remain potential sources of measurement uncertainty. To overcome these problems and to cope with the continuous increase in the number and variety of GMOs, new approaches are needed. Statistical strategies of quantification have already been proposed and expanded with the development of digital PCR. The first attempts have been made to use new generation sequencing also for quantitative purposes, although accurate quantification of the contents of GMOs using this technology is still a challenge for the future, and especially for mixed samples. New approaches are needed also for the quantification of stacks, and for potential quantification of organisms produced by new plant breeding techniques.

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Acknowledgments

We thank Dr. Christopher Berrie for reviewing the manuscript. The work was co-financed by the Slovenian Research Agency (contract no. P4-0165), the Slovenian Ministry of Agriculture and Environment (contract no. 2330-13-000072) and the Slovenian Ministry of Economic Development and Technology, Metrology Institute of the Republic of Slovenia (contract no. 640118/2008/67). The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 613908, relating to the project “Development of Cost-efficient Advanced DNA-based methods for specific Traceability issues and High Level On-site Applications”.

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Authors and Affiliations

  1. Department of Biotechnology and Systems Biology, National Institute of Biology (NIB), Večna pot 111, 1000, Ljubljana, Slovenia

    Mojca Milavec, David Dobnik, Kristina Gruden & Jana Žel

  2. Collaborative Innovation Centre for Biosafety of GMO, National Centre for Molecular Characterisation of GMOs, School of Life Science and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, China

    Litao Yang & Dabing Zhang

Authors
  1. Mojca Milavec
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  2. David Dobnik
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  3. Litao Yang
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  4. Dabing Zhang
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  5. Kristina Gruden
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  6. Jana Žel
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Corresponding author

Correspondence to Mojca Milavec.

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Published in the topical collection Nucleic Acid Quantification with guest editors Hendrik Emons and Philippe Corbisier.

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Milavec, M., Dobnik, D., Yang, L. et al. GMO quantification: valuable experience and insights for the future. Anal Bioanal Chem 406, 6485–6497 (2014). https://doi.org/10.1007/s00216-014-8077-0

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  • DOI: https://doi.org/10.1007/s00216-014-8077-0

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