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Lactococcus lactis

cells:

  1. Add 0.25 g plasmid DNA (in water) to 100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,000 V Time constant (T) 5 ms
  4. Add 1 ml ice-cold complex medium, incubate 2 hours at 30 C.
  5. Plate diluted cells on selective chloramphenicol plates. Incubate 2 days at 30 C.
Expected Results: Transformation efficiency up to 1.4 x 106 transformants/g of DNA.


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Denhardt's Solution is a mixture of blocking agents used in membrane-based hybridization protocols. The solution contains 1% Ficoll (type 400) 1% polyvinylpyrrolidone and 1% bovine serum albumin. Ste...
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