2.50
Hdl Handle:
http://hdl.handle.net/10033/616954
Title:
Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency.
Authors:
Tu, Qiang; Yin, Jia; Fu, Jun; Herrmann, Jennifer; Li, Yuezhong; Yin, Yulong; Stewart, A Francis; Müller, Rolf ( 0000-0002-1042-5665 ) ; Zhang, Youming
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.
Affiliation:
Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.
Citation:
Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency. 2016, 6:24648 Sci Rep
Journal:
Scientific reports
Issue Date:
2016
URI:
http://hdl.handle.net/10033/616954
DOI:
10.1038/srep24648
PubMed ID:
27095488
Type:
Article
Language:
en
ISSN:
2045-2322
Appears in Collections:
publications of the department of microbial natural substances ([HIPS]MINS)

Full metadata record

DC FieldValue Language
dc.contributor.authorTu, Qiangen
dc.contributor.authorYin, Jiaen
dc.contributor.authorFu, Junen
dc.contributor.authorHerrmann, Jenniferen
dc.contributor.authorLi, Yuezhongen
dc.contributor.authorYin, Yulongen
dc.contributor.authorStewart, A Francisen
dc.contributor.authorMüller, Rolfen
dc.contributor.authorZhang, Youmingen
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.en
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

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