Lactobacillus transformation (Berthier 1996)
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Revision as of 21:00, 16 September 2011
General guidelines for the electro-transformation of Lactobacillus sake as described by Berthier et al. and as used by Alegre et al. with Lactobacillus plantarum.
- 50 ul of L.plantarum competent cells
- In vitro modified plasmid DNA
- MRS media + antibiotic
- Stock solution of MgCl2
- Stock Solution of Glucose
- Electroporation Buffer (0.5M Sucrose, 10% Glycerol)
- Grow cells overnight in MRS Broth at 30°C
- Use overnight culture to inoculate 100ml MRS Broth
- Incubate at 30°C until OD600 = 0.4-0.6
All questions, input and feedback are welcome!
Berthier, F., Zagorec, M., Champomier-Verge`s, M., Ehrlich, S.D. and Morel-Deville, F. (1996) Efficient transformation of Lactobacillus sake by electroporation. Microbiology 142, 1273–1279.
Alegre et al. (FEMS Microbiology Letters 241 (2004) 73–77)
or instead, discuss this protocol. -->