Abstract
Polymerase chain reaction (PCR) is one of the most commonly used methods in diagnostic and molecular biology. However, DNA contamination can lead to incorrect results. This poses a major problem, especially in the monitoring of genetically modified organisms (GMOs). We investigated DNA contamination in 16 commercial Taq DNA polymerase reagents. Many of the Taq DNA polymerase reagents were contaminated with the 16S rRNA gene and an ampicillin resistance gene (bla). We attempted to decontaminate the Taq DNA polymerase reagents using UV, γ-ray, and electron-ray irradiation, as well as nylon membrane and FTA card treatment. UV irradiation reduced the activity of Taq DNA polymerase. Various wavelengths of γ-ray and electron-ray irradiation had no decontamination effect. However, 24-h treatment with three nylon membrane disks removed DNA contamination. FTA card treatment, which has membrane-like functions, also removed decontamination but decreased enzyme activity. We further confirmed that the nylon membrane-treated Taq DNA polymerase reagent could be used in the detection of the bla and NPTII genes in genetically modified E. coli. These results are useful for future monitoring of GMOs.
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Acknowledgements
This work was supported by the Technology Innovation Program (20014752, Development of industrial LMO monitoring technology in production and utilization facilities) funded By the Ministry of Trade, Industry & Energy (MOTIE, Korea) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2019R1I1A3A01062252).
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Zhao, S.R., Qu, B., Kim, J.K. et al. Decontamination of DNA in Taq DNA polymerase reagents using nylon membranes for monitoring of GMOs. Plant Biotechnol Rep 15, 783–790 (2021). https://doi.org/10.1007/s11816-021-00723-z
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DOI: https://doi.org/10.1007/s11816-021-00723-z