Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform.

Published

Journal Article

Electrowetting-on-dielectric (EWD) digital microfluidic laboratory-on-a-chip platforms demonstrate excellent performance in automating labor-intensive protocols. When coupled with an on-chip electroporation capability, these systems hold promise for streamlining cumbersome processes such as multiplex automated genome engineering (MAGE). We integrated a single Ti:Au electroporation electrode into an otherwise standard parallel-plate EWD geometry to enable high-efficiency transformation of Escherichia coli with reporter plasmid DNA in a 200 nL droplet. Test devices exhibited robust operation with more than 10 transformation experiments performed per device without cross-contamination or failure. Despite intrinsic electric-field nonuniformity present in the EP/EWD device, the peak on-chip transformation efficiency was measured to be 8.6 ± 1.0 × 108 cfu·μg-1 for an average applied electric field strength of 2.25 ± 0.50 kV·mm-1. Cell survival and transformation fractions at this electroporation pulse strength were found to be 1.5 ± 0.3 and 2.3 ± 0.1%, respectively. Our work expands the EWD toolkit to include on-chip microbial electroporation and opens the possibility of scaling advanced genome engineering methods, like MAGE, into the submicroliter regime.

Full Text

Duke Authors

Cited Authors

  • Madison, AC; Royal, MW; Vigneault, F; Chen, L; Griffin, PB; Horowitz, M; Church, GM; Fair, RB

Published Date

  • September 2017

Published In

Volume / Issue

  • 6 / 9

Start / End Page

  • 1701 - 1709

PubMed ID

  • 28569062

Pubmed Central ID

  • 28569062

Electronic International Standard Serial Number (EISSN)

  • 2161-5063

International Standard Serial Number (ISSN)

  • 2161-5063

Digital Object Identifier (DOI)

  • 10.1021/acssynbio.7b00007

Language

  • eng