Team:EPF-Lausanne/Protocol/Transformation

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Protocol: E.Coli Transformation


  1. Thaw the competent E.coli (DH5alpha) cells on ice (not in hands!)
  2. As soon as it is thawed, add 50µl of the cells to the DNA (~50-100 ng of pure plasmid, or some 2 µl usually)
  3. Let it rest on ice for 20-30 min. Meanwhile, put agar plate (with correct antibiotic) at 37°C for prewarming.
  4. Put the tube with DNA+E.coli at 42°C for 45 sec - 1 min (heat shock)
  5. Add 400 µl of LB broth and place at 37°C for 20-30 min (shaking)
  6. Spread the cells on the prewarmed plate (and let it dry)
  7. Incubate the plate upside-down at 37°C for ~14-15 hours (leaving it more than 16h decreases the plasmid quality)