The Xanthophyll Cycle in Photosystem II

Friday, 30 May 2014

Analysis of results, from day 1 experiments

So this Friday we basically just went in and analysed our results from FPLC the other day, having not had time the other day~

So we found a mysterious curve at the end of our results~ which indicates that there a lot of protein in our final sample where we have solublised the photosynthetic membrane.
Despite that~ we have quite a large amount of photosystem II (PSII, which I will now refer to), which is the fraction we really need for the next step of analysing on HPLC.

We came to the conclusion, that next time we want less free pigment trimmers, indicated on the graph before, as we want more of the PSII.

The graph produced via FPLC. My analysis notes in my lab book! I'll talk about my lab book in another blog :)

Therefore next time when we repeat the experiment: we will use

·        A greater sample loop for injecting into the sample loop. This is because our fractions came out extremely pale and didn’t have much green at all. If we concentrate our fractions by having a greater volume, it means that next time our fractions will be a greener colour. We increased the sample loop of the FLPC from 100ul to 500ul. However, the use of 500ul might result in a poorer resolution in our results as 500ul is the maximum volume that the sample loop can hold.

·        There is a chance that our buffer could have over solubilised our supercomplexes, giving us more free pigments than expected. Hence next time we repeat this experiment, we can try changing the concentration of the buffet- alphaDM to 0.8% instead of using 1%. This will hopefully give us a larger amount of PSII complexes obtained in the fractions next time J

I find the analysis of results the best part of experiment. I enjoy seeing outcome after a long day of slaving in the lab. I can’t wait to repeat this experiment and see what changing one variable will have an effect on the results! Also, the next time I do this experiment I will be overall more confident and know what each step is~ hence be more efficient in the lab!

Due to the fact my summer project is supervised with the lab's technician, Petra, my entire timetabling depends around her timetable. For example I do not need to come in next monday, instead I can do extra reading. This means that I'm not in everyday at the lab~ some days I actually have off to do summer related things, like going out to see friends :D Which just makes me feel even more luckier to be awarded with such studentship, as the project itself is independent- its a project of my own and not a project or another phd student. Also, its a nice summer studentship as its not 9-5pm everyday, giving me the time to actually enjoy summer! Win Win Win!

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