title: Validation of a fully integrated platform and disposable microfluidic chips enabling parallel purification of genome segments for assembly.
abstract: Recent progress in the field of genetic engineering has opened up the door to novel synthetic biology applications. Microfluidic technology has been emphasized as a key technology to support the development of these applications. While several important synthetic biology protocols have been developed in microfluidic format, no study has yet demonstrated on-chip error control. In synthetic biology protocols, the purification phase is a critical error control process which enhances the reliability of the genome segment assembly by removing undesired oligos. In this context, we report the design and characterization of a fully integrated platform, demonstrating the purification of up to 4 genome segments in parallel, prior to their off-chip assembly. The key innovation of this platform is the decoupling control strategy which eliminates the need to integrate expensive components onto the microfluidic device, enabling lower cost, disposability and rapid operation. Unlike most microfluidic chips where fluid connector plugs are needed to connect external pumps, this approach is plug-less and the chips are simply connected to the control breadboard by clamping. Furthermore the passive chip is isolated from the active control layer thereby eliminating the risk of sample-to-sample contamination in the reusable parts. As a validation of this fully-integrated system, the parallel on-chip purification of genome segments was demonstrated with ratio of correct phenotypes after final assembly up to 20% superior to the bench controls, proving thereby the suitability of the platform for synthetic biology applications.
authors.affiliation: Institute of Biological Chemistry, Biochemistry and Bioengineering, Heriot-Watt University, Edinburgh, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Kersaudy-Kerhoas
authors.forename: F
authors.initials: F
authors.lastname: Amalou
authors.forename: A
authors.initials: A
authors.lastname: Che
authors.forename: J
authors.initials: J
authors.lastname: Kelly
authors.forename: Y
authors.initials: Y
authors.lastname: Liu
authors.forename: M P Y
authors.initials: MP
authors.lastname: Desmulliez
authors.forename: W
authors.initials: W
authors.lastname: Shu
mesh_terms: D020650:Combinatorial Chemistry Techniques / Q000295:instrumentation; D004247:DNA / Q000138:chemical synthesis* / Q000302:isolation & purification; D004867:Equipment Design; D016678:Genome; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results
chemical_list: D004247:DNA
keywords: lab-on-a-chip; microfluidics; purification; synthetic biology