Week 7: I’ve Been a Busy Bee!
Hi everyone!
It has been a very busy week in the lab with a variety of different tasks! On Monday I started by finishing my last plate which included my December and January samples. I was very excited to finish the data collection part of my project (or so I thought) …. When my mentor and I were reviewing the data from all of my plates we noticed that my samples from June and July had VERY high absorbance values. Their absorbance values were above 1.o, whereas all of my other samples had values in the range of about 0.4-0.8. It’s not good to have an absorbance value above 1.0 because it could mean your samples are too concentrated and this could cause inaccuracies in measurements by the spectrophotometer. My mentor said it is entirely possible that pollens collected in the summer do just have much higher protein concentrations, however, to make sure she is going to have me run two more plates next week. The first plate will be the exact same samples again. If the absorbance values are still above 1.0, then I will have to run a second plate where we will dilute the June and July samples even further. We would want to dilute the samples so that we can have all the data fit on a straight-line curve making for easier calculations!
Speaking of calculations, I spent a lot of time on excel this week. Since I’ve never used excel before my mentor started by giving me a couple of YouTube videos to watch about basic excel commands and how to create a graph for the Bradford protein assay. My mentor also helped walk me through doing the calculations for an entire plate of samples. The math is actually a lot more complicated than I expected because there are three different tubes of samples I have to think about and convert between. The first tubes are the original 4 mL tubes that contained only pollen. The second tubes are the smaller 1.7 mL tubes that contained 1350 mL of pollen and 150 mL of NaOH. Finally, the third tubes are my 15:1 diluted samples. Sometimes when I’m doing the math I get really confused about which “tube” of sample I’m supposed to be thinking about! Luckily though, my mentor helped me set up a nice and organized chart to follow so I don’t get too lost. I forgot to take a photo, but I’ll include a picture of one of my data tables next week. Once I got the hang of excel, I was able to finish the calculations for four of my plates! Woohoo!
Additionally, this week I also worked on rewriting the “Problems” section of my project proposal packet. My mentor helped me brainstorm LOTS of potential errors that could be made during a Bradford protein assay. This included not breaking the pollen grains up enough in the bead beater, reading the samples at an incorrect wavelength, interfering substances within the pollen unintentionally reacting with Coomassie, pipetting errors, and contamination due to outside sources. Talking about all the room for error definitely made me realize how important it is that I redo the June and July samples!
I’m writing this blog a little bit earlier this week since I leave for St. Louis tomorrow to attend WashU’s admitted students’ day. I’m very excited! I hope everyone has a great week and thank you for reading!
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