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.
Highlights
The new work reported here further integrates manually generated PR-PR scripts with microfluidics to maneuver the controller of the repurposed microfluidic device to construct of DNA using a j5 [16]-designed Golden Gate assembly method [17,18,19] protocol
The script Promoter1BCD1_GFP.pr controls the input and output components by opening valves 1, 3, and 5 and closing the others so that reagents from the opened valves can flow through the ring mixer for incubation to achieve onchip DNA assembly and this assembly reaction is sent on to the output valve where the DNA assembly is collected for off-chip transformation
We used the TeselaGen DESIGN module to visually design the 4 variant combinatorial plasmid DNA library, with a common vector backbone, two promoter variants, and two Repurposing a microfluidic formulation device for automated DNA construction bicistronic device (BCD) variants coupled with a gfp gene (Fig 3)
Summary
The new work reported here further integrates manually generated PR-PR scripts with microfluidics to maneuver the controller of the repurposed microfluidic device to construct of DNA using a j5 [16]-designed Golden Gate assembly method [17,18,19] protocol. The Golden Gate DNA assembly protocol by j5 was translated manually to a PR-PR script for execution of the DNA assembly reactions on the microfluidic device.
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