UnCovid: A versatile, low-cost, and open-source protocol for SARS-CoV-2 RNA detection

Roberto Alcántara, Katherin Peñaranda, Gabriel Mendoza-Rojas, Jose A Nakamoto, Eva Dueñas, Daniela Alvarez, Vanessa Adaui, Pohl Milón

Research output: Contribution to journalArticlepeer-review

Abstract

Here, we describe a detailed step-by-step protocol to detect SARS-CoV-2 RNA using RT-PCR-mediated amplification and CRISPR/Cas-based visualization. The optimized assay uses basic molecular biology equipment such as conventional thermocyclers and transilluminators for qualitative detection. Alternatively, a fluorescence plate reader can be used for quantitative measurements. The protocol detects two regions of the SARS-CoV-2 genome in addition to the human RNaseP sample control. Aiming to reach remote regions, this work was developed to use the portable molecular workstation from BentoLab. For complete details on the use and execution of this protocol, please refer to Alcántara et al., 2021.

Original languageEnglish
Article number100878
JournalSTAR Protocols
Volume2
Issue number4
DOIs
Publication statusPublished - 2021 Dec 17
Externally publishedYes

Bibliographical note

© 2021 The Author(s).

Subject classification (UKÄ)

  • Cell and Molecular Biology
  • Infectious Medicine

Free keywords

  • COVID-19/diagnosis
  • CRISPR-Cas Systems
  • Humans
  • Nucleic Acid Amplification Techniques/methods
  • RNA, Viral/analysis
  • SARS-CoV-2/genetics

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