As noted last class, unwanted yeast growth did occur in the well plate, however, not in the expected wells. In row E, wells E7, E8, and E9 were contaminated, hence why for pH of 6.0 we will only examine two replicates per strain instead of three. In addition to this, rows D and G also experienced some contamination in wells D6 and G3.
We decided not to start the experiment all over again and went ahead to cross out the contaminated plates instead. An updated graph of the well plate set up is illustrated below.
In the presence of a sterilizing flame, 4 uL of each strain (wild-strain, SAK1-deficient strain, and TOS3-deficient strain) were inserted into their respective wells. The well plate will be placed in the spectrophotometer to measure growth rate for the next 24 hours.
Unfortunately, because the spectrophotometer had some mechanical problems, our yeast will be refrigerated to slow growth in the critical beginning phases of our data collection. As soon as the spectrophotometer is running our yeast will begin its expected growth period.
Thankfully, a working spectrophotometer was found within the next hour and our plate was able to be placed in it. The spectrophotometer was set to absorbance at 600 nanometers and continuous shake. It ran for 24hours and absorbance was measured every 20 minutes. Hopefully we get some good results next class!