TY - JOUR
T1 - PlasmidMaker is a versatile, automated, and high throughput end-to-end platform for plasmid construction
AU - Enghiad, Behnam
AU - Xue, Pu
AU - Singh, Nilmani
AU - Boob, Aashutosh Girish
AU - Shi, Chengyou
AU - Petrov, Vassily Andrew
AU - Liu, Roy
AU - Peri, Siddhartha Suryanarayana
AU - Lane, Stephan Thomas
AU - Gaither, Emily Danielle
AU - Zhao, Huimin
N1 - Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85130189437&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85130189437&partnerID=8YFLogxK
U2 - 10.1038/s41467-022-30355-y
DO - 10.1038/s41467-022-30355-y
M3 - Article
C2 - 35577775
AN - SCOPUS:85130189437
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 2697
ER -