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

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

doi:10.1038/s41467-022-30355-y

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

Journal Article · Mon May 16 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-30355-y· OSTI ID:1867945

Enghiad, Behnam; ; ; Boob, Aashutosh Girish; ; Petrov, Vassily Andrew; Liu, Roy; Peri, Siddhartha Suryanarayana; Lane, Stephan Thomas; Gaither, Emily Danielle;

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 ( Pf Ago)-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.

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Research Organization:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois at Urbana-Champaign, IL (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018420

OSTI ID:: 1867945

Alternate ID(s):: OSTI ID: 1904381
OSTI ID: 1993847

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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