Koshland Web Site/Methods
DROPOUT PLATES (3L)
DROP OUT PLATES (-ura or -leu) 3 liters
  1. 1950ml H2O
    60g agar
    --in 4L flask with stir bar
  2. 750ml H2O
    4.3g Difco yeast nitrogen base (w/o amino acids and ammonium sulfate)
    15g ammonium sulfate
    4.2g dropout powder (homemade)
  3. Dissolve in 2L flask, dissolve before autoclaving
  4. 300ml H2O
    60g glucose
    --in 1L flask, dissolve before autoclaving
  5. Autoclave 30 minutes
  6. After autoclaving equilibrate to 60-65oC in H2O bath
  7. Dry the flasks off well before mixing and pouring plates to avoid contamination
  8. When glucose solution is cooled down add:
    30ml tryptophan (10mg/ml)
    6ml threonine (20mg/ml)
  9. Mix all solutions together after cooling down. Add nutrient and glucose solutions to agar on stir plate.
Comments: Sticky notes: Bio101 YNB w/o ammonium sulfate; use 5.1g YNB and 15g ammonium sulfate;

Started supplementing B101 Dropouts with 30 ml of Ade (1mg/ml) (4/3/00)
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