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

Tu, Qiang
; 
Yin, Jia
; 
Fu, Jun
; 
Herrmann, Jennifer
; 
Li, Yuezhong
; 
Yin, Yulong
; 
Stewart, A Francis
; ; 
Zhang, Youming
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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.
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Journal
Scientific reports
URI
http://hdl.handle.net/10033/616954
DOI
10.1038/srep24648
PubMed ID
27095488
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Article
Language
en
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ISSN
2045-2322
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publications of the research group of microbial natural substances([HIPS]MINS)