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

Min Ki Shin; Seon Min Jeon; Jong Seok Park

doi:10.1007/s10068-023-01392-0

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

Food Sci Biotechnol. 2023 Aug 11;33(4):991-998. doi: 10.1007/s10068-023-01392-0. eCollection 2024 Mar.

Authors

Min Ki Shin^{1

2}, Seon Min Jeon², Jong Seok Park²

Affiliations

¹ Interdisciplinary Program in Bioengineering, Seoul National University, Seoul, 08826 Republic of Korea.
² Food Safety Risk Assessment Department, National Institute of Food and Drug Safety Evaluation, Cheongju, 28159 Republic of Korea.

PMID: 38371688
PMCID: PMC10866821 (available on 2024-08-11)
DOI: 10.1007/s10068-023-01392-0

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.

Supplementary information: The online version contains supplementary material available at 10.1007/s10068-023-01392-0.

Keywords: Conversion factor; Event-specific detection; Plasmid DNA calibrator; Quantification of GM soybean; Reference material.

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