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Homologous Recombinatorial Cloning Without the Creation of Single-Stranded Ends: Exonuclease and Ligation-Independent Cloning (ELIC)

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Abstract

We describe a new type of molecular cloning that complements the available strategies for homologous recombinatorial cloning. Purified, linear double-stranded DNA molecules with homologous ends are simply mixed in water and they transform readily into E. coli. Insert and linear vector need as few as ten base pairs of homologous sequence at their ends and essentially no incubation or enzyme treatments are needed for creating recombinants from linear fragments. Our method outcompetes most existing cloning methods in simplicity and affordability and is well-suited for high-throughput applications.

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Acknowledgments

Prof. Lloyd Ruddock and Prof. Linda M. Hendershot are kindly acknowledged for providing DNA constructs.

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Authors and Affiliations

  1. Department of Chemical Technology and Biotechnology, Aalto University, 00076, Aalto, Finland

    Essi V. Koskela & Alexander D. Frey

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  1. Essi V. Koskela
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  2. Alexander D. Frey
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Correspondence to Alexander D. Frey.

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Koskela, E.V., Frey, A.D. Homologous Recombinatorial Cloning Without the Creation of Single-Stranded Ends: Exonuclease and Ligation-Independent Cloning (ELIC). Mol Biotechnol 57, 233–240 (2015). https://doi.org/10.1007/s12033-014-9817-2

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  • DOI: https://doi.org/10.1007/s12033-014-9817-2

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