Issue 3, 2017

High efficiency hydrodynamic bacterial electrotransformation

Abstract

Synthetic biology holds great potential for addressing pressing challenges for mankind and our planet. One technical challenge in tapping into the full potential of synthetic biology is the low efficiency and low throughput of genetic transformation for many types of cells. In this paper, we discuss a novel microfluidic system for improving bacterial electrotransformation efficiency and throughput. Our microfluidic system is comprised of non-uniform constrictions in microchannels to facilitate high electric fields with relatively small applied voltages to induce electroporation. Additionally, the microfluidic device has regions of low electric field to assist in electrophoretic transport of nucleic acids into the cells. The device features hydrodynamically controlled electric fields that allow cells to experience a time dependent electric field that is otherwise difficult to achieve using standard electronics. Results suggest that transformation efficiency can be increased by ∼4×, while throughput can increase by 100–1000× compared to traditional electroporation cuvettes. This work will enable high-throughput and high efficiency genetic transformation of microbes, facilitating accelerated development of genetically engineered organisms.

Graphical abstract: High efficiency hydrodynamic bacterial electrotransformation

Supplementary files

Article information

Article type
Paper
Submitted
21 Oct 2016
Accepted
06 Dec 2016
First published
09 Jan 2017

Lab Chip, 2017,17, 490-500

High efficiency hydrodynamic bacterial electrotransformation

P. A. Garcia, Z. Ge, L. E. Kelley, S. J. Holcomb and C. R. Buie, Lab Chip, 2017, 17, 490 DOI: 10.1039/C6LC01309K

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements