Abstract
High-throughput nucleic acid extraction is critical for the implementation of modern viroid detection assays. Successful large-scale nursery, field surveys, and other regulatory, quarantine, or research diagnostic programs are increasingly dependent on high-throughput tissue pulverization and nucleic acid extraction protocols. Magnetic bead-based approaches using semi-automated robotic equipment allow high-throughput extraction and purification of high-quality uniform total nucleic acids for each individual sample. Here, we describe a streamlined and optimized protocol for citrus tissue processing and RNA extraction that can be used for downstream applications such as viroid detection by reverse transcription-quantitative polymerase chain reaction.
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Acknowledgments
Support was provided by the United States Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA), Hatch (project 1020106), and the National Clean Plant Network (NCPN) which operates under the auspices of USDA Animal and Plant Health Inspection Service (APHIS) (12-8100-1544-CA; 14-, 15-, 16-8130-0419-CA; AP17PPQS&T00C118; AP18PPQS&T00C107). The findings and conclusions in this publication are those of the author(s) and should not be construed to represent any official USDA or US government determination or policy.
URLs to sponsors’ websites are listed below.
1. CRB: https://www.citrusresearch.org
2. USDA NIFA: https://nifa.usda.gov
3. NCPN: http://nationalcleanplantnetwork.org.
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Dang, T. et al. (2022). High-Throughput RNA Extraction from Citrus Tissues for the Detection of Viroids. In: Rao, A.L.N., Lavagi-Craddock, I., Vidalakis, G. (eds) Viroids. Methods in Molecular Biology, vol 2316. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1464-8_5
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DOI: https://doi.org/10.1007/978-1-0716-1464-8_5
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1464-8
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