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Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing.
Kimbley, Lucy M; Parker, Rachel; Jongen, Maaike Sybil; Holloway, John W; Swindle, Emily J; Rose-Zerilli, Matthew Jj.
Afiliação
  • Kimbley LM; Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, UK.
  • Parker R; Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, UK.
  • Jongen MS; Centre for Hybrid Biodevices, Institute of Life Sciences, University of Southampton, Southampton, SO17 1BJ, UK.
  • Holloway JW; Human Development & Health, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, UK.
  • Swindle EJ; Human Development & Health, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, UK.
  • Rose-Zerilli MJ; Clinical & Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, UK.
Biotechniques ; 2021 Jul 16.
Article em En | MEDLINE | ID: mdl-34269076
ABSTRACT
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.
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Texto completo: 1 Bases de dados: MEDLINE Idioma: En Revista: Biotechniques Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Reino Unido

Texto completo: 1 Bases de dados: MEDLINE Idioma: En Revista: Biotechniques Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Reino Unido