Tanvi Dange, Entry #2, Plots on Plots on Plots

For the week of July 18-22, I did multiple things such as conducting another AFST on S. cerevisiae and shadow Lizzie on the flow cytometer, but the most engaging task this week was learning how to make a Job's plot. The purpose of the Job's plot is to determine the reaction stoichiometry for the formation of a metal ion complex between a metal and a ligand. For this week's Job's plots, I was studying the stoichiometry of Copper and Flu- TSCZ, which is one of Lizzie's analogues.

For the first Job's plot for Cu and Flu-TSCZ, we decided to make the highest concentration of metal/ligand 15 uM. That plot ended up looking excellent, but the concentration's were too low for drawing any conclusions, so the next day Lizzie and I decided to try again at higher concentrations.

This is what a Job's Plot should roughly look like: a pointy  mountain

The next day Lizzie and I decided to make the highest concentrations 60 uM in hopes of some data that we could use. But unfortunately, we were just having one of those days where nothing goes right. The point that was suppose to be the peak of our plot (The 5:5) just wouldn't fit the trend of our graph. We tried everything: we made multiple samples, made multiple blanks and switched machines but the point still didn't fit the data. There were still many other things to do in the lab that day, so we decided we would remake all the samples the next day and try again.

notice that one of these points is not like the others...

Finally on Friday, we scanned our new 60uM samples in the UV-vis machine and made a much more normal looking Job's plot. The Job's Plots taught me a very important lesson this week: science involves a lot of trial and error, and while it might seem frustrating to, for example, make the same plot over and over again, it's important to have patience because all of the work you do is important for the project, and the lab, as a whole.