Today we finalized our procedure to go through with our experiment as well as discovered what a FACS (Fluorescent Activated Cell Sorting) is and what it does. Note, we do not plan on using FACS because both mutations have the same color fluorescent proteins and thus will be more difficult to measure differences between them.
What we plan on doing is creating a 20 mL stock solution of YPD growth media. From there, we will split the growth media into 8 separate containers to buffer pH to have 2.o mL of solution for pHs in the range of 2.0-9.o, with an increment of 1.0 between each. We will separate each pH solution by adding 600 uL into each of 24 test tubes (one for each different strain of yeast at a different pH). We will l0ad a 96 well plate with 130 uL of each solution as the chart below shows:
There will be 4 replicates of each strain in their respective pH level. They will grow for 48 hours at 25 degrees Celsius. We will observe the growth of the cells through measuring the population density thanks to a light spectrometer, which will measure it every 15 minutes. From here, we analyze the data collected and go from there.
Here’s the link to how we are creating our buffers:
Also, we presented our third presentation: https://docs.google.com/presentation/d/1ONHKq8CehsFPntJscHWXoZZepZe4JNHQ67-DUZZdSJM/edit?usp=sharing