Team:Mississippi State/YPD Plate

From 2008.igem.org

Preparing YPD Plates

  1. Prepare 20% Dextrose solution by weighing out 40g Dextrose. Pour into dry beaker. Also get an autoclaved 500ml bottle.
  2. Add 50ml water to beaker, stirring with stirbar.
  3. When dissolved, pour contents into graduated cylinder.
  4. Add water until volume is 200ml.
  5. Pour contents into 500ml Bottle.
  6. Get another beaker.
  7. Add 2.5g yeast extract, 5g Peptone, and 200ml H2O.
  8. Stir using stirbar.
  9. When dissolved, pour contents into graduated cylinder.
  10. Add H2O until volume is 225ml.
  11. Weigh out 5g Agar, and pour into empty, autoclaved flask.
  12. Pour contents of graduated cylinder into flask, covering with foil.
  13. Take both Dextrose solution and Yeast/Peptone/Agar solution to autoclave: 20min, liquid exhaust.
  14. Go to fume hood on 3rd floor and turn on UV light to sterilize area.
  15. When autoclave finishes, immediately swirl Yeast/Peptone/Agar solution insuring everything is well dissolved.
  16. Go to fume hood on 3rd floor.
  17. Turn off UV light, and turn on normal light.
  18. Place Bottle and flask into hood.
  19. Using sterile graduated cylinder, measure out 25ml 20% Dextrose solution.
  20. Add solution to Yeast/Peptone/Agar solution to get 250ml YPD media.
  21. Allow to cool to 55C in waterbath.
  22. Pour plates (7).
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