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

Methods Mol Biol. 2022:2316:57-64. doi: 10.1007/978-1-0716-1464-8_5.

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.

Keywords: Automation; Magnetic bead extraction; Purified total RNA.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Citrus* / genetics
  • Nucleic Acids
  • RNA
  • RNA, Viral / genetics
  • Viroids* / genetics

Substances

  • Nucleic Acids
  • RNA, Viral
  • RNA