Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Qiang Tu - (Autor:in)
  • Jia Yin - , Technische Universität Dresden (Autor:in)
  • Jun Fu - (Autor:in)
  • Jennifer Herrmann - (Autor:in)
  • Yuezhong Li - (Autor:in)
  • Yulong Yin - (Autor:in)
  • A. Francis Stewart - , Professur für Biotechnologische Genomik (Autor:in)
  • Rolf Müller - (Autor:in)
  • Youming Zhang - (Autor:in)

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

OriginalspracheEnglisch
Aufsatznummer24648
FachzeitschriftScientific reports
Jahrgang6
PublikationsstatusVeröffentlicht - 20 Apr. 2016
Peer-Review-StatusJa

Externe IDs

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

Schlagworte

ASJC Scopus Sachgebiete