High-Throughput RNA Extraction from Citrus Tissues for the Detection of Viroids

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 nuclei...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2316; p. 57
Main Authors Dang, Tyler, Osman, Fatima, Wang, Jinbo, Rucker, Tavia, Bodaghi, Sohrab, Tan, Shih-Hua, Pagliaccia, Deborah, Lavagi-Craddock, Irene, Vidalakis, Georgios
Format Journal Article
LanguageEnglish
Published United States 2022
Subjects
Citrus - genetics
Nucleic Acids
RNA
RNA, Viral - genetics
Viroids - genetics
Purified total RNA
Automation
Magnetic bead extraction
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-0716-1464-8_5