PlasmidMaker is a versatile, automated, and high throughput end-to-end platform for plasmid construction

Behnam Enghiad, Pu Xue, Nilmani Singh, Aashutosh Girish Boob, Chengyou Shi, Vassily Andrew Petrov, Roy Liu, Siddhartha Suryanarayana Peri, Stephan Thomas Lane, Emily Danielle Gaither and Huimin Zhao ()
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Behnam Enghiad: University of Illinois Urbana-Champaign
Pu Xue: University of Illinois Urbana-Champaign
Nilmani Singh: University of Illinois Urbana-Champaign
Aashutosh Girish Boob: University of Illinois Urbana-Champaign
Chengyou Shi: University of Illinois Urbana-Champaign
Vassily Andrew Petrov: University of Illinois Urbana-Champaign
Roy Liu: University of Illinois Urbana-Champaign
Siddhartha Suryanarayana Peri: University of Illinois Urbana-Champaign
Stephan Thomas Lane: University of Illinois Urbana-Champaign
Emily Danielle Gaither: University of Illinois Urbana-Champaign
Huimin Zhao: University of Illinois Urbana-Champaign

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: 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. PlasmidMaker should greatly expand the potential of synthetic biology.

Date: 2022
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DOI: 10.1038/s41467-022-30355-y

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