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bioRxiv · 10.1101/2021.12.31.474679

PlasmidMaker: a Versatile, Automated, and High Throughput End-to-End Platform for Plasmid Construction

Enghiad, B.·Xue, P.·Singh, N.·Boob, A. G.·Shi, C.·Petrov, V. A.·Liu, R.·Peri, S. S.·Lane, S. T.·Gaither, E. D.·Zhao, H.
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Abstract

Plasmids are used extensively in basic and applied biology. However, design and construction of plasmids, specifically the ones carrying complex genetic information, remains one of the most time-consuming, labor-intensive, and rate-limiting steps in performing sophisticated biological experiments. Here, we report the development of a versatile, robust, automated end-to-end platform named PlasmidMaker that allows error-free construction of plasmids with virtually any sequences in a high-throughput manner. This platform consists of a most versatile DNA assembly method using Pyrococcus furiosus Argonaute (PfAgo)-based artificial restriction enzymes, a user-friendly frontend for plasmid design, and a backend that streamlines the workflow and integration with a robotic system. As a proof of concept, we used this platform to generate 101 plasmids from six different species ranging from 5 to 18 kb in size from up to 11 DNA fragments within 3 days. PlasmidMaker should greatly expand the potential of synthetic biology.

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Enghiad, B., Xue, P., Singh, N., Boob, A. G., Shi, C., Petrov, V. A., Liu, R., Peri, S. S., Lane, S. T., Gaither, E. D., Zhao, H.. 2022-01-02. PlasmidMaker: a Versatile, Automated, and High Throughput End-to-End Platform for Plasmid Construction. https://doi.org/10.1101/2021.12.31.474679

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