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Staphylococcus aureus

umes. 3. Resuspend to final volume of 1-5 x 1010 cells/ml in 10% glycerol. Aliquote and freeze at 70 C.

Electroporation of cells:

  1. Thaw cells for 5 min at room temperature. Add 1-2 l plasmid DNA (100-500 ng) to 50 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Incubate at room temperature for 30 minutes.
  2. Transfer mixture into a cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,800 V Time constant (T) 5 ms
  4. Add 1 ml SMMP50 medium to the cells and transfer to sterile tube. Incubate 90 minutes at 37 C with moderate shaking.
  5. Plate on selective BM plates. Incubate 20 hours at 37 C.
Expected Results: Transformation efficiency up to 1.5 x 105 transformants/g of DNA.


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