Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency

• DOI: 10.1038/srep24648
• Publication Date: 2016-04-20T09:40:10Z
• Authors: Qiang Tu; Jia Yin; Jun Fu; Jennifer Herrmann; Yuezhong Li; Yulong Yin; A. Francis Stewart; Rolf Müller; Youming Zhang
• Keywords: Recombineering

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 efficiency. Previous methods based on washing electroporating bacterial ice-cold condition that make them fragile prone death. Here we present simple temperature shift improve recombineering efficiency E. coli several other gram-negative bacteria thereby economizing time cost. Increased large molecules a significant advantage might facilitate cloning fragments from genomic preparations metagenomics samples.