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M-TUBE: a fabrication-free microfluidic device for large-volume bacterial electroporation requiring minimal assembly

Huang, P.-H.; Chen, S.; Shiver, A. L.; Culver, R. N.; Huang, K.; Buie, C. R.

2022-01-17 bioengineering
10.1101/2022.01.14.476275 bioRxiv
Show abstract

Conventional cuvette-based and microfluidics-based bacterial electroporation approaches 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 disposable, user-friendly microfluidic electroporation device capable of processing large volume bacterial samples yet 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.

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