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

Dataset · Fri Dec 31 00:00:00 EST 2021

DOI:https://doi.org/10.5281/zenodo.5812313· OSTI ID:3014230

Enghiad, Behnam ^[1]; Xue, Pu ^[1]; Boob, Aashutosh ^[1]; Singh, Nilmani ^[2]; Shi, Chengyou ^[1]; Lane, Stephan Thomas ^[2]; Petrov, Vassily Andrew ^[2]; Liu, Roy ^[2]; Peri, Siddhartha Suryanarayana ^[2]; Gaither, Emily Danielle ^[2]; Zhao, Huimin ^[1]

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1. Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL, USA; Department of Chemical and Biomolecular Engineering, University of Illinois Urbana-Champaign, Urbana, IL, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
2. Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

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 ~100 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.

• Research Organization: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI) (CABBI); Zenodo
• Sponsoring Organization: U.S. Department of Energy (DOE)
• DOE Contract Number: SC0018420
• OSTI ID: 3014230
• Country of Publication: United States
• Language: English