Application of Digital PCR in the Analysis of Transgenic Soybean Plants

Abstract

Detection and quantification of transgenes are important in analyzing genetically modified organisms (GMOs). Quantitative polymerase chain reaction (qPCR) is commonly utilized for such purposes. However, qPCR has certain limitations in de-tecting and quantifying transgenes in GMOs, such as the need of certified reference materials, a standard curve, and possible affection by inhibitors. Therefore, alterna-tive and possibly better methods are needed. Recent advances in digital PCR tech-nologies have promised to allow accurate quantification of nucleic acids and there-fore provided another useful technique to analyze GMOs. Thermo Fisher Scientific has recently commercialized the Applied Biosystems QuantStudio 3D digital PCR system that can be used for a wide range of applications involving nucleic acids. It will be beneficial to the scientific community to show the applicability of this digi-tal PCR system in detecting and quantifying transgenes in GMOs. In the present study, the transgenes present in the Roundup® Ready Soybean (RR1, event 40-3-2) and Roundup Ready Soybean 2 (RR2, event MON89788) developed by Monsanto Corporation were analyzed by using this digital PCR system. The qPCR analysis re-sults were included for comparison. Using specifically designed TaqMan assays, as low as 1% of the RR1 or RR2 soybean material was reliably detected and quantified on the dPCR platform. Therefore, digital PCR is a sensitive and reliable method to analyze the RR transgenic soybeans, and should be another useful tool for analyzing other transgenic plants.