Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Qiang Tu - (Author)
  • Jia Yin - , TUD Dresden University of Technology (Author)
  • Jun Fu - (Author)
  • Jennifer Herrmann - (Author)
  • Yuezhong Li - (Author)
  • Yulong Yin - (Author)
  • A. Francis Stewart - , Chair of Applied Genomics (Author)
  • Rolf Müller - (Author)
  • Youming Zhang - (Author)

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.

Details

Original languageEnglish
Article number24648
JournalScientific reports
Volume6
Publication statusPublished - 20 Apr 2016
Peer-reviewedYes

External IDs

PubMed 27095488
ORCID /0000-0002-4754-1707/work/142248097

Keywords

ASJC Scopus subject areas