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Transformation of plasmid DNA into E. coli using the heat shock method
Alexandrine Froger1, James E Hall
1Department of Physiology and Biophysics, University of California, Irvine, USA. afroger@uci.edu
Journal of Visualized Experiments : Jove
|November 11, 2008
Summary
This study details the heat shock method for transforming plasmid DNA into Escherichia coli (E. coli) bacteria. The protocol ensures successful bacterial transformation for molecular biology applications.
Area of Science:
- Molecular Biology
- Microbiology
Background:
- Plasmid DNA transformation into E. coli is a fundamental molecular biology technique.
- The heat shock method is a common approach for introducing foreign DNA into bacteria.
Purpose of the Study:
- To describe a traditional heat shock protocol for bacterial transformation.
- To demonstrate the use of commercially available chemically competent bacteria for transformation.
Main Methods:
- Chemically competent E. coli were incubated on ice with plasmid DNA.
- The mixture underwent a 45-second heat shock at 42°C, followed by re-incubation on ice.
- Transformed cells were cultured in SOC media at 37°C with agitation and plated.
Main Results:
- The traditional heat shock protocol was successfully applied to transform E. coli.
- Plating two quantities of transformed bacteria ensured colony isolation regardless of transformation efficiency.
- Commercially available competent bacteria, including Genlantis turbocells, can be used.
Conclusions:
- The described traditional heat shock protocol is effective for transforming most commercially available competent bacteria.
- Alternative rapid protocols, such as the 3-minute transformation, may be available for specific bacterial strains.
