Repurposing a microfluidic formulation device for automated DNA construction

PLoS One. 2020 Nov 11;15(11):e0242157. doi: 10.1371/journal.pone.0242157. eCollection 2020.

Abstract

Microfluidic applications have expanded greatly over the past decade. For the most part, however, each microfluidics platform is developed with a specific task in mind, rather than as a general-purpose device with a wide-range of functionality. Here, we show how a microfluidic system, originally developed to investigate protein phase behavior, can be modified and repurposed for another application, namely DNA construction. We added new programable controllers to direct the flow of reagents across the chip. We designed the assembly of a combinatorial Golden Gate DNA library using TeselaGen DESIGN software and used the repurposed microfluidics platform to assemble the designed library from off-chip prepared DNA assembly pieces. Further experiments verified the sequences and function of the on-chip assembled DNA constructs.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computational Biology
  • DNA / analysis*
  • Escherichia coli / metabolism
  • Lab-On-A-Chip Devices*
  • Microfluidics / instrumentation*
  • Microfluidics / methods*
  • Oligonucleotide Array Sequence Analysis*
  • Pattern Recognition, Automated
  • Polymerase Chain Reaction
  • Software

Substances

  • DNA

Grants and funding

The study was supported by NSF SBIR grant IIP-1430986 and the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research. Work conducted at TeselaGen Biotechnology by MF and NE was funded by NSF SBIR. Work conducted at JBEI by GG, NJH, and GL was funded by the DOE Joint BioEnergy Institute (https://www.jbei.org) supported by the U. S. Department of Energy, Office of Science, Office of Biological and Environmental Research, through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the U.S. Department of Energy. The funding agencies provided support in the form of labor and supplies costs for authors GG, NE, MF, NJH, and GL. The funding agencies did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. We declare that this work was done in a collaboration with TeselaGen Biotechnology, which is a commercial entity.