Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing
Autor: | Lucy M Kimbley, Rachel Parker, Maaike Sybil Jongen, John W Holloway, Emily J Swindle, Matthew JJ Rose-Zerilli |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: | |
Zdroj: | BioTechniques, Vol 71, Iss 1, Pp 387-391 (2021) |
Druh dokumentu: | article |
ISSN: | 1940-9818 0736-6205 |
DOI: | 10.2144/btn-2021-0020 |
Popis: | Single-cell RNA sequencing (scRNA-seq) of the bronchial epithelium enables examination of cellular subtypes and their responses to viral infections. Here, an optimized method for the isolation of virally infected primary bronchial epithelial cells using a commercially available microfluidic device is presented. Using this method single cells can be rapidly isolated with minimal equipment available in most laboratories. Isolation can be carried out inside biological safety cabinets, permitting the use of virally infected cells. Both cell-line and primary cells isolated using the device retained sufficient RNA integrity for the generation of short-read sequencing-compatible cDNA libraries to facilitate scRNA-seq. |
Databáze: | Directory of Open Access Journals |
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