RT Journal Article
SR Electronic
T1 Antha-guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2019.12.17.879486
DO 10.1101/2019.12.17.879486
A1 P. Handal Marquez
A1 M. Koch
A1 D. Kestemont
A1 S. Arangundy-Franklin
A1 V. B. Pinheiro
YR 2019
UL http://biorxiv.org/content/early/2019/12/18/2019.12.17.879486.abstract
AB Protein engineering through directed evolution facilitates the screening and characterization of protein libraries. Efficient and effective methods for multiple site-saturation mutagenesis, such as Darwin Assembly, can accelerate the sampling of relevant sequence space and the identification of variants with desired functionalities. Here, we present the automation of the Darwin Assembly method, using a Gilson PIPETMAX™ liquid handling platform under the control of the Antha software platform, which resulted in the accelerated construction of complex, multiplexed gene libraries with minimal hands-on time and error-free, while maintaining flexibility over experimental parameters through a graphical user interface rather than requiring user-driven library-dependent programming of the liquid handling platform. We also present an approach for barcoding libraries that overcomes amplicon length limitations in next generation sequencing and enables fast reconstruction of library reads.