A Versatile Microfluidic Device for Automating Synthetic Biology

作者：Shih Steve C C; Goyal Garima; Kim Peter W; Koutsoubelis Nicolas; Keasling Jay D; Adams Paul D; Hillson Nathan J; Singh Anup K

来源：ACS Synthetic Biology, 2015, 4(10): 1151-1164.

DOI：10.1021/acssynbio.5b00062

• digital microfluidics
• droplet microfluidics
• synthetic biology
• DNA assembly
• Golden Gate assembly
• Gibson assembly
• yeast assembly
• TAR cloning

New microbes are being engineered that contain the genetic circuitry, metabolic pathways, and other cellular functions required for a wide range of applications such as producing biofuels, biobased chemicals, and pharmaceuticals. Although currently available tools are useful in improving the synthetic biology process, further improvements in physical automation would help to lower the barrier of entry into this field. We present an innovative microfluidic platform for assembling DNA fragments with 10x lower volumes (compared to that of current microfluidic platforms) and with integrated region-specific temperature control and on-chip transformation. Integration of these steps minimizes the loss of reagents and products compared to that with conventional methods, which require multiple pipetting steps. For assembling DNA fragments, we implemented three commonly used DNA assembly protocols on our microfluidic device: Golden Gate assembly, Gibson assembly, and yeast assembly (i.e., TAR cloning, DNA Assembler). We demonstrate the utility of these methods by assembling two combinatorial libraries of 16 plasmids each. Each DNA plasmid is transformed into Escherichia coli or Saccharomyces cerevisiae using on-chip electroporation and further sequenced to verify the assembly. We anticipate that this platform will enable new research that can integrate this automated microfluidic platform to generate large combinatorial libraries of plasmids and will help to expedite the overall synthetic biology process.

• 出版日期2015-10