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Actinomyces viscosus

poration of cells:
  1. Thaw bacterial suspensions on ice. Add up to 5 l (100 ng) plasmid DNA to 100 l (5 x 108) 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) 2,500 V Time constant (T) 5 ms
  4. Dilute cells with 1 ml of CAM and incubate for 2 h at 37 C.
  5. Plate onto selective brain heart infusion agar plates; incubate at 37 C for up to 5 days.

Expected results:

Transformation efficiency up to 2.9 x 107 transformants/g of DNA.


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