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Bifidobacterium animalis

of 1 mM ice-cold sucrose-citrate buffer, dispense in tubes and incubate for 3.5 hours at 4 C.

Electroporation of cells:

  1. Add 0.5-1.5 g plasmid DNA to 80 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) 1,200 V Time constant (T) 5 ms
  4. Dilute with 800 l outgrowth medium and incubate for 2.5 h at 37 C.
  5. Plate onto selective MRS agar plates; incubate anaerobically for 2-3 days at 37 C.
Expected Results: Transformation efficiency up to 9.4 x 104 transformants/g of DNA.


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