Experiment: In this activity yo Page 5 ze the results of experiments that investigate nutritional requirement of several mutant strains of yeast. The mutations in these strains cause a nutritional requirement for an amino acid, such that the strains will not grow in media that lack one specific amino acid. Any mutant that has a nutritional requirement is called an auxotroph, and is incapable of growing in a "minimal medium" containing only a carbon source (e.g., glucose), a simple nitrogen source (e.g., ammonium sulfate), and various salts and minerals. Such strains can be supported on a medium supplemented with only the missing nutrient or on a "rich" medium that contains amino acids, vitamins, nitrogenous bases, etc. (often in the form of an extract from yeast). The wild-type individual that can synthesize the metabolic component is a prototroph, and is capable of growth on minimal medium. The mutant strains in this activity are unable to synthesize tryptophan, lysine, or histidine; one prototrophic strain is included in the analysis. We start with a master plate that has 7 haploid strains of yeast (A-G), arranged in patches in the following order (looking at the plate right side up with the upper surface of the agar facing you): A B с D E F G The yeast strains from the master plate will be tested for their ability to grow on five different media. A very light inoculum of each strain will be transferred in the same pattern as above to the following media: unsupplemented minimal medium, three minimal media each supplemented by one amino acid (tryptophan, histidine, or lysine), and a rich medium. After inoculation, the plates are incubated at 30°C for one or two days, and grow or absence of growth is recorded. Question 6: What do you think is the composition of the medium in the master plate (minimal, minimal supplemented with one amino acid, or rich)? O Rich O Minimal supplemented with one amino acid O Minimal

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