Team:Imperial College London/Wetlab/Protocols/LB Plates

From 2009.igem.org



LB Plates Protocol

Makes 8 plates

  1. Take a clean conical flask and add 200ul distilled water from the dispenser.
  2. Weigh out the correct mass of LB Agar according to the packet instructions, and add to the conical flask with the water. Put in foam bung, cover with foil, tape with autoclave tape, and send to the autoclave for sterilisation.
  3. Once the agar has been sterilised in the autoclave, remove of the foil and remelt the agar mixture in the microwave until the mixture boils.
  4. When the mixture has completely dissolved, add to a preheated waterbath at 50'C to allow the temperatures to equilibrate.
  5. Add 200ul antibiotic to the flask when the temperature is suitable, and mix by swirling.
  6. Pour the mixture between 8 plastic plates.
  7. Using a bunsen burner, wave the flame over the plates to remove any air bubbles and to sterilise.
  8. Allow to cool underneath a Bunsen flame, to keep the plates sterile.
  9. Once dry, turn the plates over, secure with parafilm and store the cold room until needed.


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