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Quantitative real-time PCR analysis of four genetically modified soybean events using plasmid and genomic DNA calibrators

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Abstract

A calibration curve is required for reliable and accurate quantitative real-time PCR analysis of genetically modified (GM) organisms, necessitating the use of reference materials as calibrators. In this study, two types of DNA calibrators—plasmid DNA (pDNA) and genomic DNA (gDNA)—were used to quantify four GM soybean events (SYHT0H2, MON87751, DAS-44406-6, and DAS-81419-2). The PCR efficiency and linearity for the calibrators adhered to the CODEX guidelines. The conversion factor (Cf) was calculated as the ratio of copies of GM events to those of endogenous genes using the pDNA calibration curve. To assess the accuracy and repeatability of these assays, quantification at GM levels of 3% and 1% was performed. Based on our results, we believe that the pDNA calibrator assessed in this study can be used as a reference material for GMO quantitative analysis and can replace gDNA, especially considering the ease of management and advantages of mass production.

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Acknowledgements

This study was supported by the Ministry of Food and Drug Safety of Korea.

Funding

This study was funded by the Ministry of Food and Drug Safety of Korea (Grant Number: 19161MFDS052).

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Authors and Affiliations

  1. Interdisciplinary Program in Bioengineering, Seoul National University, Seoul, 08826, Republic of Korea

    Min Ki Shin

  2. Food Safety Risk Assessment Department, National Institute of Food and Drug Safety Evaluation, Cheongju, 28159, Republic of Korea

    Min Ki Shin, Seon Min Jeon & Jong Seok Park

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  1. Min Ki Shin
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  2. Seon Min Jeon
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  3. Jong Seok Park
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MKS and SMJ contributed equally to this work.

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Correspondence to Min Ki Shin.

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Shin, M.K., Jeon, S.M. & Park, J.S. Quantitative real-time PCR analysis of four genetically modified soybean events using plasmid and genomic DNA calibrators. Food Sci Biotechnol 33, 991–998 (2024). https://doi.org/10.1007/s10068-023-01392-0

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