Yeast metabolic footprinting (YMF) medium
Glucose, 2% (w/v) (Sigma-Aldrich G5767)
H2O, ultrapure (MilliQ or Barnstead)
Metabolite cocktail, filter-sterilized, each component at 1mM final concentration
Yeast nitrogen base without amino acids and ammonium sulfate, 0.17% (w/v) (BD 233520)
Ammonium sulfate, 0.5% (w/v)
METHOD
Basic Preparation
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1. To prepare YMF medium for culture in 250-mL flasks, combine the glucose, ultrapure H2O, yeast nitrogen base and ammonium sulfate.
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2. Dispense 41 mL of this mixture into each flask.
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3. Autoclave at 121ºC for 15 min.
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4. Allow the medium to cool, and add 9 mL of filter-sterilized metabolite cocktail to each flask.
Large-Scale Preparation
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1. To ensure uniformity, autoclave 1-L volumes of the glucose-yeast nitrogen base-ammonium sulfate mixture and pool into a single, large container.
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2. Add 180 mL of filter-sterilized metabolite cocktail per liter, and aseptically dispense 50 mL of the supplemented medium per 250-mL sterile flask.
YMF medium is a synthetic, defined, minimal medium that is supplemented with a metabolite cocktail of amino acids, organic acids, and pyrimidine bases. It was designed to minimize the interference of small inorganic ions with mass spectrometric analyses.
