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Related Experiment Video

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The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection
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Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform.

Andrew C Madison1, Matthew W Royal1, Frederic Vigneault2

  • 1Department of Electrical and Computer Engineering, Duke University , Durham, North Carolina 27708, United States.

ACS Synthetic Biology
|June 2, 2017
PubMed
Summary

This study integrates electrowetting-on-dielectric (EWD) digital microfluidics with on-chip electroporation for efficient bacterial transformation. This advances automated genome engineering (MAGE) in small volumes.

Keywords:
digital microfluidicsdropletelectroporationtransformation

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Area of Science:

  • Microfluidics
  • Molecular Biology
  • Bioengineering

Background:

  • Electrowetting-on-dielectric (EWD) digital microfluidic platforms automate complex laboratory protocols.
  • Integrating electroporation with EWD systems can streamline genome engineering processes like multiplex automated genome engineering (MAGE).

Purpose of the Study:

  • To integrate an electroporation electrode into an EWD platform for high-efficiency bacterial transformation.
  • To demonstrate the feasibility of on-chip microbial electroporation within a digital microfluidic system.

Main Methods:

  • A Ti:Au electroporation electrode was incorporated into a parallel-plate EWD device.
  • Escherichia coli transformation was performed using reporter plasmid DNA in 200 nL droplets.
  • Device performance was evaluated for robustness, cross-contamination, and transformation efficiency.

Main Results:

  • The integrated EWD-electroporation device achieved robust operation with over 10 experiments per device without failure.
  • Peak on-chip transformation efficiency reached 8.6 ± 1.0 × 108 cfu·μg-1 at 2.25 ± 0.50 kV·mm-1 electric field.
  • Cell survival and transformation fractions were 1.5 ± 0.3% and 2.3 ± 0.1%, respectively.

Conclusions:

  • The EWD toolkit now includes on-chip microbial electroporation capabilities.
  • This integration enables scaling of advanced genome engineering methods, such as MAGE, to submicroliter volumes.