Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/13/2/167 |
| _version_ | 1827343005073276928 |
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| author | Gail L. Sturdevant Kimberly D. Meade-White Sonja M. Best Emily Speranza |
| author_facet | Gail L. Sturdevant Kimberly D. Meade-White Sonja M. Best Emily Speranza |
| author_sort | Gail L. Sturdevant |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-07T22:18:40Z |
| format | Article |
| id | doaj.art-5b877b4717a34e428dcc232105ab2f33 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-07T22:18:40Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-5b877b4717a34e428dcc232105ab2f332024-02-23T15:30:25ZengMDPI AGPathogens2076-08172024-02-0113216710.3390/pathogens13020167Preservation of scRNA-Seq Libraries Using Existing Inactivation ProtocolsGail L. Sturdevant0Kimberly D. Meade-White1Sonja M. Best2Emily Speranza3Innate Immunity and Pathogenesis Section, Laboratory of Neurological Infections and Immunity, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USADisease Modeling and Transmission Section, Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USAInnate Immunity and Pathogenesis Section, Laboratory of Neurological Infections and Immunity, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USAFlorida Research and Innovation Center, Cleveland Clinic Lerner Research Institute, Port Saint Lucie, FL 34987, USASingle-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.https://www.mdpi.com/2076-0817/13/2/167single-cell RNA sequencingbiosafety level 4sample inactivation |
| spellingShingle | Gail L. Sturdevant Kimberly D. Meade-White Sonja M. Best Emily Speranza Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols Pathogens single-cell RNA sequencing biosafety level 4 sample inactivation |
| title | Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols |
| title_full | Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols |
| title_fullStr | Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols |
| title_full_unstemmed | Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols |
| title_short | Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols |
| title_sort | preservation of scrna seq libraries using existing inactivation protocols |
| topic | single-cell RNA sequencing biosafety level 4 sample inactivation |
| url | https://www.mdpi.com/2076-0817/13/2/167 |
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