Actinobacillus Pleuropneumoniae

Actinobacillus pleuropneumoniae

Making electrocompetent cells:

1. Grow cells to mid-exponential growth phase at an O.D.₆₅₀of 0.5.
2. Harvest by centrifugation at 3,000 x g for 10 min at 4 °C.
3. Wash twice in 15% glycerine at 4 °C.
4. Resuspend in 1/20 volume of 15% glycerine and keep at 4 °C.

Electroporation of cells:

1. Add 3 µl (300 µg/ml) plasmid DNA to 125 µl of electrocompetent cells. Homogenize by gently mixing with pipette
several times. Transfer mixture into a prechilled cuvette.
2. Wipe moisture from the cuvette and insert the cuvette into the device.
3. Electroporation:

Expected results:

Transformation efficiency up to 1 x 10⁷ transformants/µg of DNA.