Repurposing a microfluidic formulation device for automated DNA construction

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.

High-throughput structure determination of an intrinsically disordered protein using cell-free protein crystallization

“…The trial numbers of crystallization are expected to be improved by the automated system for DNA construction and protein expression in the future. (54)(55)(56)(57) An automated screening system can be applied for comprehensive analysis, such as an alanine scan, to establish the crystal design strategy. Crystal design will be diversified by using composite crystals of proteins and organic compounds as scaffold crystals.…”

BioCloneBot: A versatile, low-cost, and open-source automated liquid handler

“…Consequently, do-it-yourself (DIY) biologists and engineers have taken on the task of developing affordable, open-source hardware to democratize automation. Currently, a multitude of open-source DIY solutions are available, encompassing syringe pumps [8] , [9] , [10] , peristaltic pumps [11] , plate-readers [12] , microfluidic devices [13] , [14] , [15] , [16] , [17] , [18] and a wide array of other innovations.…”

Automation and Expansion of EMMA Assembly for Fast-Tracking Mammalian System Engineering

“…Through the use of a bottom-up approach, individual genetic parts, or functional modules, are characterized independently and then combined to build novel and complex genetic circuits. , Recently, the introduction of standardized modular platforms for the design and assembly of mammalian gene expression cassettes has allowed the systematic dissection and quantitation of the function of diverse DNA regulatory modules in high throughput and at high sensitivity. − A comprehensive review of mammalian genetic toolkits was recently provided by Di Blasi et al Libraries of vectors provide powerful tools for biological investigation, most notably for massively parallel interrogation of protein and genetic sequence function and other cell-based assays. However, despite the availability of multiple modular assembly platforms and the increasing application of automation to DNA assembly, − there remains no robust, well-characterized system for the automated production of large arrayed libraries of mammalian expression vectors in a laboratory setting. Without access to a flexible system for generating vector libraries for parallel testing, the ability of researchers to leverage the extensive power of mammalian synthetic biology at high throughput is severely limited.…”