M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform.

Autor: Po-Hsun Huang, Sijie Chen, Anthony L Shiver, Rebecca Neal Culver, Kerwyn Casey Huang, Cullen R Buie
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: PLoS Biology, Vol 20, Iss 9, p e3001727 (2022)
Druh dokumentu: article
ISSN: 1544-9173
1545-7885
DOI: 10.1371/journal.pbio.3001727
Popis: Conventional cuvette-based and microfluidics-based electroporation approaches for bacterial gene delivery have distinct advantages, but they are typically limited to relatively small sample volumes, reducing their utility for applications requiring high throughput such as the generation of mutant libraries. Here, we present a scalable, large-scale bacterial gene delivery approach enabled by a disposable, user-friendly microfluidic electroporation device requiring minimal device fabrication and straightforward operation. We demonstrate that the proposed device can outperform conventional cuvettes in a range of situations, including across Escherichia coli strains with a range of electroporation efficiencies, and we use its large-volume bacterial electroporation capability to generate a library of transposon mutants in the anaerobic gut commensal Bifidobacterium longum.
Databáze: Directory of Open Access Journals
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