06/15/2015

New Microfluidics DNA Assembly Platform

The Science

Microbes are being engineered for a wide range of applications such as producing biofuels, biobased chemicals, and pharmaceuticals. Although currently available tools are useful for this process, further improvements are needed to lower the barriers scientists face if they plan to enter this growing field. Researchers at the Department of Energy’s Joint BioEnergy Institute have developed an innovative microfluidic platform for assembling DNA fragments, a critical step in the entire process. The new system uses volumes 10 times lower than current microfluidic platforms and has integrated region-specific temperature control and on-chip transformation. Integration of these steps in a single device minimizes the loss of reagents and products compared to conventional methods, which require, for example, multiple pipetting steps. For assembling DNA fragments, researchers implemented three commonly used DNA assembly protocols on the new microfluidic device: Golden Gate assembly, Gibson assembly, and yeast assembly (i.e., TAR cloning, DNA Assembler). Assembly of two combinatorial libraries of 16 plasmids each demonstrated the utility of these microfluidic methods. Each DNA plasmid was transformed into Escherichia coli or Saccharomyces cerevisiae using on-chip electroporation and further sequenced to verify the assembly. This platform likely will enable new research that can integrate this automated microfluidic platform to generate large combinatorial libraries of plasmids, helping to expedite the overall synthetic biology process for biofuels development.

Principal Investigator

Anup K. Singh
Sandia National Laboratories

Co-Principal Investigator

Steve C. C. Shih
Lawrence Berkeley National Laboratory

BER Program Manager

Shing Kwok

U.S. Department of Energy, Biological and Environmental Research (SC-33)
Biological Systems Science Division
[email protected]

References

Shih, S. C. C., G. Goyal, P. W. Kim, N. Koutsoubelis, J. D. Keasling, P. D. Adams, N. J. Hillson, and A. K. Singh. 2015. “A Versatile Microfluidic Device for Automating Synthetic Biology,” ACS Synthetic Biology. DOI: 10.1021/acssynbio.5b00062.