Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols

Author:

Sturdevant Gail L.1,Meade-White Kimberly D.2,Best Sonja M.1,Speranza Emily3

Affiliation:

1. Innate Immunity and Pathogenesis Section, Laboratory of Neurological Infections and Immunity, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA

2. Disease Modeling and Transmission Section, Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA

3. Florida Research and Innovation Center, Cleveland Clinic Lerner Research Institute, Port Saint Lucie, FL 34987, USA

Abstract

Single-cell RNA sequencing has soared in popularity in recent years. The ability to deeply profile the states of individual cells during the course of disease or infection has helped to expand our knowledge of coordinated responses. However, significant challenges arise when performing this analysis in high containment settings such as biosafety level 3 (BSL-3), BSL-3+ and BSL-4. Working in containment is necessary for many important pathogens, such as Ebola virus, Marburg virus, Lassa virus, Nipah and Hendra viruses. Since standard operating procedures (SOPs) for inactivation are extensive and may compromise sample integrity, we tested whether the removal of single-cell sequencing libraries from containment laboratories using existing inactivation protocols for nucleic acid extraction (Trizol, RLT buffer, or AVL buffer) was feasible. We have demonstrated that the inactivation does not affect sample quality and can work with existing methods for inactivation.

Funder

National Institute of Allergy and Infectious Diseases of the National Institutes of Health

Cleveland Clinic Foundation

Publisher

MDPI AG

Reference25 articles.

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3. (2023, November 28). Prioritizing Diseases for Research and Development in Emergency Contexts. Available online: https://www.who.int/activities/prioritizing-diseases-for-research-and-development-in-emergency-contexts/prioritizing-diseases-for-research-and-development-in-emergency-contexts.

4. (2023, November 28). HHS and USDA Select Agents and Toxins, Available online: https://www.selectagents.gov/sat/list.htm.

5. CDC (2018). Guidance on the Inactivation or Removal of Select Agents and Toxins for Future Use, 7 CFR Part 331, 9 CFR Part 121.3, 42 CFR Part 73.3.

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