Yeast Sensitivity to Caffeine
The purpose of this lab is to test the sensitivity to the caffeine of knock out strains of yeast cells. We will be analyzing the data on the gene's response to different levels of caffeine concentration.
- 10 mM powdered caffeine
- 100 mM of caffeine solution
- Wild Type Yeast
- Knock out genes
- Assay Plate
- 1.5 mL test tubes
- Incubator set at 30 degrees celsius
- Find concentrations of caffeine solution needed for experiment
- Combine caffeine solution with Wild Type yeast (BY4735) in assay plate
- Incubate at 30 degrees celsius overnight
- For our experiment we used concentration levels of .2mM, .5mM, 1mM, 2mM, 5mM, 10mM, 20 mM, 50mM, and 100mM.
- We combined 10uL of each caffeine solution with 90uL of Wild Type Yeast-our control- (BY4735) in the assay tray
- After analyzing our data we chose to use our .2mM concentration to test on a variety of different yeast genes including:
*YG6235W *YJL043W *YCL002C *YMR090W *YCR100C *YPL068C *YEL035C *YFL064C (Our gene)
The data presented is our calibration experiment with wild type yeast combined with various amounts of caffeine concentration. As you can see, the yeast cells were extremely sensitive to the caffeine, and the only clear data was the .2 mM concentration level. We decided to use that concentration point to test on the knock out genes. Looking at the data, the majority of the yeast cells died at .5mM concentration level.
- Doubling time was 38.0198 minutes