SD + LYS + α-Aminoadipate Medium
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1. Dissolve 1.7 g of yeast nitrogen base without ammonium sulfate and without amino acids and 20 g of dextrose in 100 mL of dH2O. Add 2 mL of 1.5% l-lysine. Filter-sterilize.
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2. Dissolve 2 g of DL-α-aminoadipate in 100 mL of dH2O. Add pellets of KOH until all of the DL-α-aminoadipate goes into solution. Then adjust the pH to 5.5 with glacial acetic acid. Filter-sterilize.
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3. Add 20 g of Bacto agar to 800 mL of dH2O, and autoclave.
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4. Add the filter-sterilized solutions to the autoclaved agar, mix thoroughly, and pour the medium into Petri dishes.
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Although it is possible to add some nutritional supplements to α-aminoadipate medium, the best results are obtained with either no supplements (using a strain containing only the required lys5 mutation) or supplements that either cannot be utilized as nitrogen sources (e.g., adenine, histidine, uracil) or are poorly utilized (e.g., tryptophan).
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This recipe was adapted from Chattoo et al. (1979) and Ito-Harashima and McCusker (2004).
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