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USER-Derived Cloning Methods and Their Primer Design

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DNA Cloning and Assembly Methods

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1116))

Abstract

Uracil excision-based cloning through USERâ„¢ (Uracil-Specific Excision Reagent) is an efficient ligase-free cloning technique that comprises USER cloning, USER fusion, and USER cassette-free (UCF) USER fusion. These USER-derived cloning techniques enable seamless assembly of multiple DNA fragments in one construct. Though governed by a few simple rules primer design for USER-based fusion of PCR fragments can prove time-consuming for inexperienced users. The Primer Help for USER (PHUSER) software is an easy-to-use primer design tool for USER-based methods. In this chapter, we present a PHUSER software protocol for designing primers for USER-derived cloning techniques.

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Acknowledgement

This work was supported by a strategic PhD stipend from the Faculty of LIFE Sciences.

Author information

Authors and Affiliations

  1. DynaMo Center of Excellence, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Copenhagen, Denmark

    Bo Salomonsen & Barbara A. Halkier

  2. Department of Systems Biology, Center for Microbial Biotechnology, Technical University of Denmark (DTU), Kongens Lyngby, Denmark

    Uffe H. Mortensen

Authors
  1. Bo Salomonsen
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  2. Uffe H. Mortensen
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  3. Barbara A. Halkier
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Editor information

Editors and Affiliations

  1. Norwegian University of Science and Technology, Trondheim, Norway

    Svein Valla

  2. Norwegian University of Science and Technology, Trondheim, Norway

    Rahmi Lale

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Salomonsen, B., Mortensen, U.H., Halkier, B.A. (2014). USER-Derived Cloning Methods and Their Primer Design. In: Valla, S., Lale, R. (eds) DNA Cloning and Assembly Methods. Methods in Molecular Biology, vol 1116. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-764-8_5

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  • DOI: https://doi.org/10.1007/978-1-62703-764-8_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-763-1

  • Online ISBN: 978-1-62703-764-8

  • eBook Packages: Springer Protocols

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