Quick Blog update: Had meeting with Professor Ruban himself on Thursday and discovered how to further proceed with my project. He wanted me to run HPLC on my B4 fraction from BBY spinach preparation obtained from FPLC. The de epxodiation I yielded in my A9 fractions, containing PSII supercomplex, was sufficiently lower than expected, lower than that of my isolation of stacked thylakoids, which should not be the case. This means something wasn't right with the prep. He told me to check the HPLC on the B4 fractions (Where the Light harvesting II complexes were situated.) De epoxidation in B4 fractions should be 100%. However, with out HPLC analysis we found values much of 70%, therefore we clearly knew something was wrong with the preparation.
Now I know for next time: That I must always run HPLC on my B4 fractions, fractions contianing LHCII supercomplexes to see if it is worth the continuation of the BBY spinach preparation past the steps which are identical in protocol to that of isolation of stacked thylakoids.
Furthermore, the increasing the time of stacking time in the BBY medium. Allow the stacking of thylakoids allows more PSII supercomplexes to be present in the preparation as PSII is mainly found in the stacked thylakoids, compared to that of PSII found mainly in the lamellae of the chloroplast.
Anyways enough science talk, some nice pictures taken over the week
|Irina's healthy growing Peas, wow from 3 weeks they have grown so much! Check earlier blogs to see how they looked at the start! We are just putting wooden sticks in to ensure they grow upright and not tangled :) Happy gardening !|
|Supervising Irina on HPLC analysis~ teaching her the ways!|
|Here is Petra grinding up ivy leaves using liquid nitrogen. Yes the steam coming off the ivy is liquid nitrogen that Petra has poured on the leaves, freezing them, making them into crystals, easier to grind into pieces.|
|Petra adding the ivy crystals into a test tube ready for it to be grinded by hand. Everything done under iced conditions again.|