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使用现有灭活方案保存单细胞RNA测序文库

Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols.

作者信息

Sturdevant Gail L, Meade-White Kimberly D, Best Sonja M, Speranza Emily

机构信息

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.

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

出版信息

Pathogens. 2024 Feb 13;13(2):167. doi: 10.3390/pathogens13020167.

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.

摘要

近年来,单细胞RNA测序越来越受欢迎。在疾病或感染过程中深入分析单个细胞状态的能力,有助于拓展我们对协调反应的认识。然而,在生物安全3级(BSL-3)、BSL-3+和BSL-4等高防护环境中进行这种分析时,会出现重大挑战。对于许多重要病原体,如埃博拉病毒、马尔堡病毒、拉沙病毒、尼帕病毒和亨德拉病毒,在防护环境中开展工作是必要的。由于核酸提取的现有灭活标准操作程序(SOP)内容繁多,可能会损害样本完整性,因此我们测试了使用现有的核酸提取灭活方案(Trizol、RLT缓冲液或AVL缓冲液)从防护实验室中取出单细胞测序文库是否可行。我们已经证明,这种灭活不会影响样本质量,并且可以与现有的灭活方法配合使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63d7/10891800/4f34ca3bbab5/pathogens-13-00167-g001.jpg

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