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The budding yeast Saccharomyces cerevisiae has served as one of the most important model organism for the study of eukaryotic molecular biology. The entire yeast genome has been sequenced and the function of many yeast genes is now known. This rapid progress has relied heavily on the ability to introduce replicating plasmids into yeast. A typical replicating yeast plasmid contains an origin of replication known as an autonomously replicating sequence or ARS and a selectable marker gene that enables the yeast to grow in the absence of a nutrient such as uracil. In this exercise, students introduce such a plasmid into yeast and then select for transformants by plating the cells on a nutrient-agar medium lacking uracil. They then study the stability of the plasmid in yeast cells by growing the cells in the presence of uracil. The program requires basic training in microbiology and was designed for 8 groups of students. The following materials and equipment are needed but not included: microcentrifuge, table-top centrifuge, autoclave to prepare media, bacterial spreading rod, Bunsen burners, sterile water and microliter dispensers.
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