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[Bio101] A Quick and Easy Method for Making Competent Escherichia coli Cells for Transformation Using Rubidium Chloride   

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Abstract

This protocol describes a quick and efficient method to make competent E. coli cells for transformation using rubidium chloride. Commercial competent cells are expensive and this protocol provides a cheaper alternative to them.

Keywords: Competent cells, E. coli, Transformation efficiency, TOP10, DH5α

Background

The success of gene cloning is highly dependent on the transformation efficiency of bacterial cells. The efficiency can be artificially improved by treating the cells with chemicals or electric pulses. Several protocols are available to prepare competent E. coli cells, however, they are usually long, laborious, and show inconsistency in competence. The protocol by Green and Rogers (2013) overcomes these downsides and allows the preparation of highly competent cells (~106-108 CFU/µg DNA). While other protocols require cells to be grown at low temperature (19-22 °C), this protocol involves growing cells at 37 °C. Thus, the cells grow faster and reach log phase within 4 h as compared to 18-24 h. This protocol is highly reproducible.

Copyright: © 2017 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Sharma, N., Anleu Gil, M. X. and Wengier, D. (2017). A Quick and Easy Method for Making Competent Escherichia coli Cells for Transformation Using Rubidium Chloride. Bio-protocol Bio101: e2590. DOI: 10.21769/BioProtoc.2590.
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