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dc.contributor.authorTu, Qiang
dc.contributor.authorYin, Jia
dc.contributor.authorFu, Jun
dc.contributor.authorHerrmann, Jennifer
dc.contributor.authorLi, Yuezhong
dc.contributor.authorYin, Yulong
dc.contributor.authorStewart, A Francis
dc.contributor.authorMüller, Rolf
dc.contributor.authorZhang, Youming
dc.date.accessioned2016-07-14T08:29:43Z
dc.date.available2016-07-14T08:29:43Z
dc.date.issued2016
dc.identifier.citationRoom temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency. 2016, 6:24648 Sci Repen
dc.identifier.issn2045-2322
dc.identifier.pmid27095488
dc.identifier.doi10.1038/srep24648
dc.identifier.urihttp://hdl.handle.net/10033/616954
dc.description.abstractBacterial 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.
dc.language.isoenen
dc.titleRoom temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.en
dc.identifier.journalScientific reportsen
refterms.dateFOA2018-06-13T00:35:44Z
html.description.abstractBacterial 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.


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