Lactobacillus transformation (Berthier 1996)

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Overview

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.

Materials

• 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)

Procedure

1. Grow cells overnight in MRS Broth at 30°C
2. Use overnight culture to inoculate 100ml MRS Broth
3. Incubate at 30°C until OD₆₀₀ = 0.4-0.6

Notes

All questions, input and feedback are welcome!

References

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)

Contact

• morto077@uottawa.ca

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