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ZeBRα a universal, multi-fragment DNA-assembly-system with minimal hands-on time requirement

DOI zum Zitieren der Version auf EPub Bayreuth: https://doi.org/10.15495/EPub_UBT_00004650
URN to cite this document: urn:nbn:de:bvb:703-epub-4650-2

Title data

Richter, David ; Bayer, Katharina ; Toesko, Thomas ; Schuster, Stefan:
ZeBRα a universal, multi-fragment DNA-assembly-system with minimal hands-on time requirement.
In: Scientific Reports. Vol. 9 (27 February 2019) Issue 1 .
ISSN 2045-2322
DOI der Verlagsversion: https://doi.org/10.1038/s41598-019-39768-0

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Abstract

The recently evolved field of synthetic biology has revolutionized the way we think of biology as an “engineerable” discipline. The newly sprouted branch is constantly in need of simple, cost-effective and automatable DNA-assembly methods. We have developed a reliable DNA-assembly system, ZeBRα (Zero-Background Redα), for cloning multiple DNA-fragments seamlessly with very high efficiency. The hallmarks of ZeBRα are the greatly reduced hands-on time and costs and yet excellent efficiency and flexibility. ZeBRα combines a “zero-background vector” with a highly efficient in vitro recombination method. The suicide-gene in the vector acts as placeholder, and is replaced by the fragments-of-interest, ensuring the exclusive survival of the successful recombinants. Thereby the background from uncut or re-ligated vector is absent and screening for recombinant colonies is unnecessary. Multiple fragments-of-interest can be assembled into the empty vector by a recombinogenic E. coli-lysate (SLiCE) with a total time requirement of less than 48 h. We have significantly simplified the preparation of the high recombination-competent E. coli-lysate compared to the original protocol. ZeBRα is the least labor intensive among comparable state-of-the-art assembly/cloning methods without a trade-off in efficiency.

Further data

Item Type: Article in a journal
DDC Subjects: 500 Science
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology > Chair Animal Physiology > Chair Animal Physiology - Univ.-Prof. Dr. Stefan Schuster
Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology > Chair Animal Physiology
Language: English
Originates at UBT: Yes
URN: urn:nbn:de:bvb:703-epub-4650-2
Date Deposited: 27 Mar 2020 11:14
Last Modified: 27 Mar 2020 11:14
URI: https://epub.uni-bayreuth.de/id/eprint/4650

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