Electro-transformation of Lactobacillus spp.

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(New page: ==Overview== Instructions on how to prepare ''Lactobacillus plantarum'' competent cells before electrotransformation. =='''Materials'''== *MRS media *Overnight culture of L. plantarum c...)
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'''Relevant Papers and Books'''
'''Relevant Papers and Books'''
Alegre et al (FEMS Microbilogy Letters 241 (2004) 73-77)
Alegre et al (FEMS Microbiology Letters 241 (2004) 73-77)

Revision as of 13:54, 18 October 2009



Instructions on how to prepare Lactobacillus plantarum competent cells before electrotransformation.


  • MRS media
  • Overnight culture of L. plantarum cells
  • 20 ml solution of MgCL2 (10mM)
  • 20 ml solution of sucrose (0.5 M) and glycerol (10%w/v)


  1. Inoculate L. plantarum overnight using 10 ml of MRS media.
  2. The next morning inoculate 100 ml of MRS with the 10 ml overnight culture.
  3. Incubate cells for 3-4 hrs at 30°C unitl absorbance reaches 600nm=0.85.
  4. Pellet cells by centrifugation at max speed (keep cells cold on ice in between centrifugations) for approx. 2 minutes or until supernatant is clear.
  5. Wash cells twice with 10 ml of chilled MgCL2 solution.
  6. Resuspend cells in 3-4ml of sucrose/glycerol solution used to wash cells.
  7. Can then be stored in an ice bath for no more than 4hrs before use.
  8. Store in aliquots and store in a -80°C freezer.


All questions, input and feedback are welcome!

  1. Centrifugation at 4000 rpm for 2 minutes was sufficient to pellet the competent cells to give a clear supernatant.


Relevant Papers and Books

Alegre et al (FEMS Microbiology Letters 241 (2004) 73-77)


  • morto077@uottawa.ca

or instead, discuss this protocol. -->

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