Transformation of plasmid DNA into E. coli using the heat shock method.
Transformation of plasmid DNA into E. coli using the heat shock method is a basic technique of molecular biology. It consists of inserting a foreign plasmid or ligation product into bacteria. This video protocol describes the traditional method of transformation using commercially available chemically competent bacteria from Genlantis. After a short incubation in ice, a mixture of chemically competent bacteria and DNA is placed at 42 degrees C for 45 seconds (heat shock) and then placed back in ice. SOC media is added and the transformed cells are incubated at 37 degrees C for 30 min with agitation. To be assured of isolating colonies irrespective of transformation efficiency, two quantities of transformed bacteria are plated. This traditional protocol can be used successfully to transform most commercially available competent bacteria. The turbocells from Genlantis can also be used in a novel 3-minute transformation protocol, described in the instruction manual.
Froger
Alexandrine
A
Department of Physiology and Biophysics, University of California, Irvine, USA. afroger@uci.edu
Hall
James E
JE
D019411:Clinical Laboratory Techniques; D004247:DNA; D004926:Escherichia coli / Q000235:genetics*; D006358:Hot Temperature; D007053:Ice; D010957:Plasmids; D014169:Transformation, Bacterial
D007053:Ice; D004247:DNA