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    Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency.

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    Authors
    Tu, Qiang
    Yin, Jia
    Fu, Jun
    Herrmann, Jennifer
    Li, Yuezhong
    Yin, Yulong
    Stewart, A Francis
    Müller, Rolf cc
    Zhang, Youming
    Issue Date
    2016
    
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    Abstract
    Bacterial competent cells are essential for cloning, construction of DNA libraries, and mutagenesis in every molecular biology laboratory. Among various transformation methods, electroporation is found to own the best transformation efficiency. Previous electroporation methods are based on washing and electroporating the bacterial cells in ice-cold condition that make them fragile and prone to death. Here we present simple temperature shift based methods that improve DNA transformation and recombineering efficiency in E. coli and several other gram-negative bacteria thereby economizing time and cost. Increased transformation efficiency of large DNA molecules is a significant advantage that might facilitate the cloning of large fragments from genomic DNA preparations and metagenomics samples.
    Citation
    Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency. 2016, 6:24648 Sci Rep
    Affiliation
    Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.
    Journal
    Scientific reports
    URI
    http://hdl.handle.net/10033/616954
    DOI
    10.1038/srep24648
    PubMed ID
    27095488
    Type
    Article
    Language
    en
    ISSN
    2045-2322
    ae974a485f413a2113503eed53cd6c53
    10.1038/srep24648
    Scopus Count
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