Abstract
Magnetic iron oxide nanoparticles functionalized with 4-carboxyphenylboronic acid (CPBA-MNPs) were developed for extracting genomic DNA, total RNA and nucleic acids from seeds. The seed samples were genetically-modified maize seeds and unmodified soybean seeds infected by bean pod mottle virus and tobacco ringspot virus. The total nucleic acids, genomic DNA, and RNA could be separately extracted from these seeds with high qualities using CPBA-MNPs under different conditions. Furthermore, the results of real-time quantitative qPCR and real-time reverse transcription (RT)-PCR indicated that the nucleic acids extracted from these seeds using CPBA-MNPs were suitable for the detection of genetically-modified seeds and seed-borne viruses.
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Acknowledgments
This work was supported by a Special Fund from the General Administration of Quality Supervision (201110035 and 201310068) and the Chinese Ministry of Science and Technology (2011CB932803).
Supporting information
Reaction systems of real-time qPCR and RT-PCR
PCR amplification of 18S rDNA from nucleic acid extracted from soybean seed
Supplementary Table 1—Primers 1 and probes used in this study
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Ning Sun and Congliang Deng have contributed equally to this work.
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Sun, N., Deng, C., Ge, G. et al. Application of carboxyphenylboronic acid-functionalized magnetic nanoparticles for extracting nucleic acid from seeds. Biotechnol Lett 37, 211–218 (2015). https://doi.org/10.1007/s10529-014-1658-3
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DOI: https://doi.org/10.1007/s10529-014-1658-3