Multiporator / Electroporator 2510
Transformation Protocol
Protocol No. 4308 915.532 01/2002
Microorganism
Salmonella typhimurium LT2
Cell type
Bacteria, gram negative
Molecules injected
Plasmid DNA (pBR322 in TE buffer)
Growth medium
LB medium
Washing solution
10 mM HEPES, pH 7.0; 10% glycerol
Electroporation solution
10% glycerol
Outgrowth medium
SOC medium (without antibiotics)
Cuvette
2 mm gap width
Reference
Binotto J., et al 1991 Canadian Journal of Microbiology
37 474-477
Making electrocompetent cells:
1.
Incoculate a flask of LB 1:100 with a fresh overnight
culture, grow at 37 C with shaking to an O.D.
640 of 0.75. Chill
the cells in an ice-water bath for 15 min.
2.
Harvest by centrifugation (4,000 x g, 10 min., 4 C).
3.
Wash one time in the original culture volume with HEPES
and one time with 10% glycerol using 1/100 of the original volume.
4.
Resuspend in 10% glycerol.
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Salmonella typhimurium