Room Temperature Electrocompetent Bacterial Cells Improve DNA Transformation and Recombineering Efficiency

Overview

Journal Sci Rep

Specialty Science

Date 2016 Apr 21

PMID 27095488

Citations 35

Authors

Qiang Tu

Jia Yin

Jun Fu

Jennifer Herrmann

Yuezhong Li

Yulong Yin

A Francis Stewart

Rolf Muller

Youming Zhang

Affiliations

Soon will be listed here.

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.

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