Team:Mississippi State/Making LB Media for Plates

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  • Making LB Media for plates and making LB broth are exactly the same, except for the addition of Agar to plate media.
  1. For every 100ml LB to be made, get 2.5g LB powder.
  2. !!!Important!!! Only do this step if making media for plates. For every 100ml LB to be made, get 1.5g Agar.
  3. Combine LB powder and 100ml water in 250ml autoclaved flask.
  4. !!!Important!!! Only do this step if making media for plates. Add Agar.
  5. Cover with foil and autoclave tape.
  6. Swirl until dissolved.
  7. Autoclave: Fill water in autoclave to top of from lip. Close, set time to 20-25min. It will usually be 40-50min before you can open it. Make sure exhaust is set to Slow(Liquids).
  8. After 40-50min., put on heat gloves and slowly open machine. Watch out for steam!!!
  9. Remove flask, place on metallic surface until it cools.
  10. If making LB Broth, you are done at this point. Close bottle.
  11. Don't allow Media to cool too much, when warm to touch, pour into plates.
  12. Get 1 plate for every 20-25ml Media.
  13. Pour media into each plate until surface is covered.
  14. If media remains, distribute evenly among other plates.
  15. Allow plates to cool, then flip upside-down.
  16. Wait overnight.
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