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Propionibacterium freudenreichii

ut 1/100 of the original culture volume).

Electroporation of cells:

  1. Add plasmid DNA to 80-100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,000 V Time constant (T) 5 ms
  4. Immediately add 900 l cold outgrowth medium and incubate for 2.5-3 h at 30 C.
  5. Plate cells onto selective SLB plates; incubate 5-7 days at 30 C under anaerobic conditions.
Expected Results: Transformation efficiency up to 1 x 108 transformants/g of DNA.


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