Jajesniak, P. and Wong, T.S. (2016) Rapid Construction of Recombinant Plasmids by QuickStep-Cloning. Methods in Molecular Biology, 1472. pp. 205-214. ISSN 1940-6029
Abstract
QuickStep-Cloning is a novel molecular cloning technique that builds upon the concepts of asymmetric PCR and megaprimer-based amplification of whole plasmid. It was designed specifically to address the major drawbacks of previously reported cloning methods. The fully optimized protocol allows for a seamless integration of a long DNA fragment into any position within a plasmid of choice, in a time-efficient and cost-effective manner, without the need of a tedious DNA gel purification, a restriction digestion, and an enzymatic ligation. QuickStep-Cloning can be completed in less than 6 h, significantly faster than most of the existing cloning methods, while retaining high efficiency.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2016 Springer Science+Business Media New York. This is an author produced version of a paper subsequently published in Methods in Molecular Biology. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Recombinant DNA; Megaprimer PCR; Ligation-independent cloning; Plasmid construction; Synthetic biology; Protein engineering; Metabolic engineering |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Chemical and Biological Engineering (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 03 Jan 2017 11:09 |
Last Modified: | 27 Sep 2017 20:07 |
Published Version: | https://doi.org/10.1007/978-1-4939-6343-0_16 |
Status: | Published |
Publisher: | Springer Verlag |
Refereed: | Yes |
Identification Number: | 10.1007/978-1-4939-6343-0_16 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:109509 |