A highly efficient and highly reliable protocol for transformation of Escherichia coli by electroporation

Shuli Wang, Yanfeng Luo, Xiao Yi, Wenwen Yu, Zhiwu Xu, Xiaoqian Ma, Jianguo He, Qiuyun Liu

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Electroporation is widely adopted for highly efficient transformation of Escherichia coli of different purposes, but its efficiency is highly varied in the range of several orders of magnitude. Here we describe a modified and highly reliable electroporation protocol which combines low temperature growth in the presence of sucrose with a postelectroporation heat pulse, which not only gives rise to an over 250-fold enhancement in transformation compared with conventional protocols, but also alleviates the need for a strict low temperature control at all times of an electroporation-competent cell preparation experiment and highly pure MilliQ grade water which is not readily available in many laboratories. Transformations for a 6-kb E. coli and yeast shuttle vector were always higher than 107 transformants/μg, and more than 108 transformants/μg were sometimes recorded in routine experiments.

Original languageEnglish (US)
Pages (from-to)253-258
Number of pages6
JournalJournal of Rapid Methods and Automation in Microbiology
Volume15
Issue number3
DOIs
StatePublished - Sep 1 2007

Fingerprint Dive into the research topics of 'A highly efficient and highly reliable protocol for transformation of Escherichia coli by electroporation'. Together they form a unique fingerprint.

Cite this