Team:EPF-Lausanne/Protocol/AgarPlates

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Protocol: Agar Plates


  • Add to a bottle:
    • 20 g/l of LB broth powder.
    • 10 g/l of Agar (not agarose).
  • Fill up with DI water and autoclave (program 106, takes 2 hours). Remember to leave the cap loose!
  • Label the plates (found in the stock room).
  • When the autoclaving is done, close the bottle cap and take the bottle out.
  • Let rest until it cools down to around 55ºC (can be held for some seconds): if warmer, the antibiotic degrades, if colder, the broth gelifies.
  • Add the antibiotic (Ampicillin 100 µg/ml) = 1 ml of 100 mg/ml Amp for 1l of broth. Same for chloramphenicol or Spe.
  • Add 25-30 ml of broth to each plate and let open for 2 hours.
  • Close plates, wrap in alu foil (Amp is sensitive to light) and store at +4ºC.