The “Yeast Mutation and Analysis” lab protocol served as a guideline for the first day of our Yeast Mutagenesis experiment. During the first day, we performed a serial dilution of ~107to ~104yeast cells using a pipette, sterile test tubes, and vortexer. From the105dilution, we pipetted 100µL onto a YED plate and repeated with a second YED plate. We then exposed both plates to UV radiation for 5 seconds. We made a control plate that we did not expose to UV radiation and incubated all three plates for 3-5 days. After that time, the lab professor removed them from the incubator. During the next lab period on September 26th, we observed the plates. Since we had no mutants, mutated yeast cells were provided to us. We designed an experiment to test what gene the mutants were mutated in. We decided to cross the 4 unknown types of mutants we were given with 4 known types of mutants to see which mutants complemented and which mutants had mutations in the same genes. During the same lab period, we streaked the 4 unknown mutants and the 4 known mutant parent groups onto a YED plate and left them to incubate for two days. The streaks contained 1a, 2a, aMw, and aMx on the horizontal axis and 1ɑ, 2ɑ, ɑMy, and ɑMz on the vertical axis. On September 28th, two days after streaking and incubation, we mated the UV mutants. A small sample of each of the corresponding parent colonies was put where the two parent groups would intersect and mixed with each other. We allowed 2 days of incubation after mating before replica plating onto an MV plate. On October 2nd, the YED plate was replica plated onto an MV plate and an MV+Ade plate. We left the replica MV plates in the incubator until the following lab period.
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