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

View ORCID ProfilePo-Hsun Huang, Sijie Chen, View ORCID ProfileAnthony L. Shiver, Rebecca Neal Culver, Kerwyn Casey Huang, Cullen R. Buie
doi: https://doi.org/10.1101/2022.01.14.476275
Po-Hsun Huang
1Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
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Sijie Chen
1Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
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Anthony L. Shiver
2Department of Bioengineering, Stanford University, Stanford, CA 94305
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Rebecca Neal Culver
3Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305
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Kerwyn Casey Huang
2Department of Bioengineering, Stanford University, Stanford, CA 94305
4Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305
5Chan Zuckerberg Biohub, San Francisco, CA 94158
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Cullen R. Buie
1Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
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  • For correspondence: crb@mit.edu
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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.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 17, 2022.
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M-TUBE: a fabrication-free microfluidic device for large-volume bacterial electroporation requiring minimal assembly
Po-Hsun Huang, Sijie Chen, Anthony L. Shiver, Rebecca Neal Culver, Kerwyn Casey Huang, Cullen R. Buie
bioRxiv 2022.01.14.476275; doi: https://doi.org/10.1101/2022.01.14.476275
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M-TUBE: a fabrication-free microfluidic device for large-volume bacterial electroporation requiring minimal assembly
Po-Hsun Huang, Sijie Chen, Anthony L. Shiver, Rebecca Neal Culver, Kerwyn Casey Huang, Cullen R. Buie
bioRxiv 2022.01.14.476275; doi: https://doi.org/10.1101/2022.01.14.476275

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