DNA Assembly in 3D Printed Fluidics

Abstract

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.

Open Source 3D‐printed focussing mechanism for cellphone‐based cellular microscopy

“…The increasing access to 3D printers has opened up possibilities in Open Source development of scientific instrumentation in the biological context as demonstrated by the OpenLabware project (Baden et al, 2015). This has also become a part of the increasing movement in synthetic biology as evidenced by the increasing interest in the iGEM synthetic biology contest for hardware coupled to 'wet-ware', such as the OpenScope project entry in the 2015 iGEM (Cambridge-JIC 2015iGEM, 2015 and the advantages of integrating it with pumps, fluidics and cloning in a single device (Patrick et al, 2015). Recent reports of real-time PCR devices for diagnostics (Mulberry et al, 2017) suggest an integration of a focussed microscope combined with such devices, which could result in increased access to devices that combine biochemical and genetic diagnostics with optics.…”

Open-source, community-driven microfluidics with Metafluidics

“…The advent of soft lithography was a groundbreaking enabling technology Open-source, community-driven microfluidics with Metafluidics p e r s p e c t i v e for microfluidics 20 , but photolithography requirements-from clean rooms to technology for silicon processing-still present a substantial obstacle for both developers and users. Increasingly, commodity digital fabrication technologies like three-dimensional (3D) printing 21 are being used to manufacture molds for soft lithography 22 and milli-and microfluidic systems 23,24 , including programmable valves 25 and devices for synthetic biology applications 26 . Crucially, 3D printing eliminates the photolithographic step and obviates the need for expensive silicon-processing infrastructure.…”

The upcoming 3D-printing revolution in microfluidics

“…DMD-based SL has been used to create hollow, ~ 1 mm long micro-needles (with a bore diameter of 375 μm) that could penetrate cadaveric porcine skin 22 (Figure 3B). Lately, microfluidic devices for immunomagnetic separation of bacteria 23 , separation of cells by using helical channels with trapezoid cross-sections 24 (Figure 3C), gradient generation 25 (Figure 3D), emulsion droplet generators 25-28 , DNA assembly 29 and an oxygen control insert for a 24-well dish 30 , to name a few, have been designed using single-photon SL. The minimum cross-sectional area of a microchannel that is attainable by SL depends not only on the laser spot-size or pixel resolution, but also on the type and viscosity of the resin, which has to be effectively drained from the channels post-printing 31 .…”