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Mycobacterium smegmatis

Electroporation of cells:

  1. Add up to 1 g plasmid DNA to 60 l of electrocompetent cells. Homogenize by gently mixing with pipette several
    times. Incubate on ice for 10 min. Transfer mixture into a prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,200 V Time constant (T) 5 ms
  4. Wash cells out of the cuvette with 1 ml of outgrowth medium and incubate for 2 h at 37 C.
  5. Plate on selective agar.
Expected Results: Transformation efficiency up to 105 transformants/g of DNA.


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