DNA Assembly in 3D Printed Fluidics

The process of connecting genetic parts—DNA assembly—is a foundational technology for synthetic biology. Microfluidics present an attractive solution for minimizing use of costly reagents, enabling multiplexed reactions, and automating protocols by integrating multiple protocol steps. However, microfluidics fabrication and operation can be expensive and requires expertise, limiting access to the technology. With advances in commodity digital fabrication tools, it is now possible to directly print fluidic devices and supporting hardware. 3D printed micro- and millifluidic devices are inexpensive, easy to make and quick to produce. We demonstrate Golden Gate DNA assembly in 3D-printed fluidics with reaction volumes as small as 490 nL, channel widths as fine as 220 microns, and per unit part costs ranging from $0.61 to $5.71. A 3D-printed syringe pump with an accompanying programmable software interface was designed and fabricated to operate the devices. Quick turnaround and inexpensive materials allowed for rapid exploration of device parameters, demonstrating a manufacturing paradigm for designing and fabricating hardware for synthetic biology.基因元件连接——即DNA组装（DNA assembly）——是合成生物学的核心基础技术。微流控技术（microfluidics）可为该领域提供极具吸引力的解决方案：其可减少昂贵试剂的消耗、支持多重反应，并通过集成多步实验流程实现实验方案的自动化。然而，微流控器件的制备与操作往往成本高昂，且需具备专业技术能力，这制约了该技术的普及应用。随着商用数字制备工具的技术迭代，如今已可直接打印流控器件及其配套硬件。3D打印的微流控与毫流控器件成本低廉、制备简便且生产高效。本研究验证了在3D打印流控器件中开展Golden Gate DNA组装（Golden Gate DNA assembly）的可行性，其反应体积可低至490纳升，通道宽度最小可达220微米，单份元件的成本区间为0.61美元至5.71美元。研究团队还设计并制备了一款搭载可编程软件界面的3D打印注射泵，用于操控此类流控器件。得益于材料成本低廉且生产周期极短，研究团队得以快速探索器件参数，由此验证了一种适用于合成生物学硬件设计与制备的新型制造范式。