Automation of DNA processing based on DNA reuse (Leader: RUB)
Objectives
- Automation of DNA processing biochemistry using off-the-shelf liquid handling and lab automation technologies
- Develop an open-source robot programming language for automating DNA processing and DNA-based applications
- Customizing automated robotics for selected applications that test the DNA libraries
- Developing custom DNA processing micro-fluidics technology
Advanced liquide logic collaboration
Microfluidics may revolutionize our ability to write synthetic DNA by addressing several fundamental limitations associated with generating novel genetic constructs. Here we report the de novo synthesis and cell-free cloning of custom DNA libraries in sub-microliter reaction droplets using programmable digital microfluidics. Specifically, we developed Programmable Order Polymerization (POP), Microfluidic Combinatorial Assembly of DNA (M-CAD) and Microfluidic In-vitro Cloning (MIC) and applied them to de-novo synthesis, combinatorial assembly and cloning of genes, respectively. We demonstrate the utility of these methodologies in synthetic biology by programming an autonomous microfluidic system to construct and clone designer DNA libraries, which were then retrieved in individual droplets and applied to the study of yeast ribosome binding sites and bacterial Azurin. The ability to rapidly and robustly generate designer DNA molecules in a completely programmable and autonomous manner should have
wide application in biological research and development.
