PlasmidMaker

Enghiad, B., Xue, P., Boob, A., Singh, N., Shi, C., Lane, S.T., Petrov, V.A., Liu, R., Peri, S.S., Gaither, E.D., Zhao, H. Dec. 31, 2021. “PlasmidMaker: A Versatile, Automated, and High Throughput End-to-End Platform for Plasmid Construction.” Zenodo. DOI: 10.5281/zenodo.5812313.

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 three days. PlasmidMaker should greatly expand the potential of synthetic biology.

PlasmidMaker Web Application

Frontend and Backend Code

Download (7.3 KB) includes:

• 101 Plasmid DNA Sequences (.csv format)
• Primer and Guide DNA Sequences (.dna format)
• API Endpoint Guide