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Development and applications of real-time PCR standards for GMO quantification based on tandem-marker plasmids

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An Erratum to this article was published on 19 October 2005

Abstract

The need to comply with mandatory thresholds introduced for the labeling of GM food and feed by recent European Regulations is increasing the interest for reliable standards for GMO quantification. Certified reference materials (CRMs) for the different GMO lines are an essential element to calibrate measurement systems adopted to quantify GM ingredients. Unfortunately, conventional CRMs are affected not only by their high production costs but also by time consuming production and certification procedures. At the moment CRMs are available only for few of the GMOs authorized in Europe. In addition, latest developments in European legislation makes conceivable that the number of different authorized GM lines to be analysed will increase significantly, raising the need for additional CRMs. In the field of GMO detection and quantification, plasmids have been demonstrated to represent a cheap and reliable alternative to conventional CRMs. Here we describe the production of tandem-marker plasmids containing simultaneously specific target sequences for the GMO of interest and a species-specific reference marker sequence in a 1:1 ratio have been developed and used as real-time PCR standards. Strengths and weaknesses of these novel standards are discussed in detail.

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Acknowledgements

E. M. and C. B. did all the cloning and real-time PCR laboratory work; F. W. ideated and developed the technique, the working strategy, elaborated data and wrote the manuscript. M. Q. provided as laboratory supervisor all the scientific and logistical support needed for the experiments and G. V.d.E. is the head of the Biotechnology and GMOs Unit of the JRC. Mr. M. Maretti was helpful in his lab technician duties. Finally, the authors whish to thank Dr. S. Langrell who provided a critical reading of the manuscript

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Authors and Affiliations

  1. European Commission, Joint Research Centre (JRC), Institute for Health and Consumer Protection (IHCP), Biotechnology & GMOs Unit, Via Enrico Fermi, 1, I-21020, Ispra, VA, Italy

    Elia Mattarucchi, Florian Weighardt, Cristina Barbati, Maddalena Querci & Guy Van den Eede

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  1. Elia Mattarucchi
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  2. Florian Weighardt
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  3. Cristina Barbati
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  4. Maddalena Querci
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  5. Guy Van den Eede
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Corresponding author

Correspondence to Maddalena Querci.

Additional information

An Erratum for this chapter can be found at http://dx.doi.org/10.1007/s00217-005-0148-1

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Mattarucchi, E., Weighardt, F., Barbati, C. et al. Development and applications of real-time PCR standards for GMO quantification based on tandem-marker plasmids. Eur Food Res Technol 221, 511–519 (2005). https://doi.org/10.1007/s00217-005-1168-6

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  • DOI: https://doi.org/10.1007/s00217-005-1168-6

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