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Development of certified matrix-based reference material of genetically modified rice event TT51-1 for real-time PCR quantification

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Abstract

In 2009, the genetically modified (GM) rice event TT51-1 with an engineered insect resistance trait became the first GM rice event to be granted certification for safe production in China, and its derivative lines Bt 63 and Huahui No.1 are expected to be commercialized soon. The development of certified reference material (CRM) for TT51-1 is necessary to monitor and inspect the TT51-1 event and its derivates. In this work, we developed four matrix-based TT51-1 rice CRMs (TT51-1a, TT51-1b, TT51-1c, and TT51-1d) with different TT51-1 mass fraction ratios by blending seed powders of homozygous TT51-1 and its recipient cultivar Minghui 63. The between-bottle homogeneity and the within-bottle homogeneity were tested, and good results were obtained. The potential degradation during transportation and shelf life were evaluated, and demonstrated an expiration period of at least 36 months. The characterization values of the four TT51-1 CRMs based on the mass fraction ratio were 1000.000 ± 51.430 g/kg, 49.940 ± 4.620 g/kg, 9.990 ± 1.110 g/kg, and 4.990 ± 0.620 g/kg, respectively. The characterization values based on the copy number ratio were certified by digital PCR analysis as 97.442 ± 5.253 %, 4.851 ± 0.486 %, 1.042 ± 0.135 %, and 0.556 ± 0.073 %, respectively. These results suggested that the TT51-1 matrix-based CRMs developed are of high quality and can be used as potential calibrators for TT51-1 GM rice inspection and monitoring.

The general procedure for the development of TT51-1 rice certified reference materials

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Acknowledgments

This work was supported by the National Transgenic Plant Special Fund (2014ZX08012-003), the National Natural Science Foundation of China (31471670), and te Program for New Century Excellent Talents in University.

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Authors and Affiliations

  1. National Center for the Molecular Characterization of Genetically Modified Organisms, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Yu Jiang, Hui Yang, Sheng Quan, Yinan Liu & Litao Yang

  2. Development Center of Science and Technology, Ministry of Agriculture of People’s Republic of China, Beijing, 100025, China

    Ping Shen

Authors
  1. Yu Jiang
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  2. Hui Yang
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  3. Sheng Quan
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  4. Yinan Liu
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Corresponding author

Correspondence to Litao Yang.

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Jiang, Y., Yang, H., Quan, S. et al. Development of certified matrix-based reference material of genetically modified rice event TT51-1 for real-time PCR quantification. Anal Bioanal Chem 407, 6731–6739 (2015). https://doi.org/10.1007/s00216-015-8836-6

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  • DOI: https://doi.org/10.1007/s00216-015-8836-6

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