Growth of yeast cells on solid media to test for functionality of endogenous or heterologous ammonium transporters.
Method Article
Yeast growth assays and quantification
https://doi.org/10.1038/nprot.2007.86
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Growth of yeast cells on solid media to test for functionality of endogenous or heterologous ammonium transporters.
Transform and select yeast on solid YNB (minimal yeast medium without nitrogen) medium supplemented with 3% glucose and 1 mM arginine. For selection, grow cells in liquid YNB medium supplemented with 3% glucose and 1 mM arginine for one day, dilute 10, 100, 1000 and 10000 in water and drop 5 µL of cells on solid YNB medium containing 50 mM Tris/MES buffered at pH 5.2, 3% glucose and 0.2, 0.4 or 2 mM ammonium chloride or, as a control, 1 mM arginine.
Incubate cells 3-6 days at 28ºC and document results by
scanning the plates at 600dpi in grey scale.
This protocol has been posted on Protocol Exchange, an open repository of community-contributed protocols sponsored by Nature Portfolio. These protocols are posted directly on the Protocol Exchange by authors and are made freely available to the scientific community for use and comment.
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