Methods of S.cerevisiae Complementation

The first step of this experiment was setting up matings of different strains in patches on a YED plate(a rich medium that contains all the nutrients yeast need to grow). The mating types that were crossed included HB1xHA0, HB1xHA1, HB1xHA2, and HA1xHA2. In order to carry out these crosses, a toothpick was used to transfer the appropriate haploid strain onto the plate by simply using a sterile toothpick to transfer a dollop of the mating partner on top and gently mixing the two strains on the surface of the agar. The steps were repeated for the remaining crosses and the plates were left incubated at 30° C. YED plates contain adenine so even the haploid adenine deficient cells were able to grow, so in order to determine which crosses were able to grow without the presence of adenine, the cells were placed on MV plates as well(media that contains minimum amount of nutrients required to support growth). Both the haploid and diploid yeast strains were plated into quadrants of MV plates using the same techniques that were used on the YED plates.