Well, it’s now Thursday and this whole week has absolutely flown by!
I won’t go through this in too much detail and won’t separate the days because, as with last week, it’s kind of been pretty samey each day. A lot of what I’ve been doing (almost all of it) has been lab based and important to my project, and I don’t want to reveal too much incase I do end up putting my report on here.!
This week I’ve been working on my main experiment for my project. I began the cell stimulations on Thursday (as mentioned before) and began the Western Blotting on Monday.
Unfortunately, I was actually supposed to finish the practical on Tuesday afternoon but after 2 whole days of really hard work, I found that one of my bands hadn’t shown up! I was absolutely devastated actually, because I’d put so much effort in!
It turned out that one of the doctors who was demonstrating had given me some bad advice, which resulted in me cutting through the band, rather than around it, and so when it came to adding the antibody, it was only able to attach right at the edge of the membrane, making it pretty much indistinguishable, and completely unquantifiable when it came to scanning and analysis.
So yes, I was gutted. And then I had to spend yesterday and today completely redoing the Western Blot from scratch. Weirdly, I don’t mind that much actually. I really enjoy being in the lab and so I’m happy to do it again. It also meant that I was able to get more practical experience while here, and I think that I learnt quite a lot doing it again a bit more independently.
I had various people helping me out the final time because nobody had two days completely free. It made it perhaps more confusing as everybody has a different technique for the same process, and so I was often being told a lot of conflicting information that I was reassured was the best way of doing it… haha! Overall, I had much less help than before, and was able to take a bit more responsibility for my work. This I enjoyed.
Regarding the first experiment… On the bright side, the band that did show up (pERK) was absolutely beautiful! It was exactly as my model had predicted and so I was able to make some pretty awesome figures showing the data, and will have a lot to talk about in my report.
In the end, I was able to get some results. This was a big relief after effectively 4 days of solid Western Blotting. The results weren’t as pure and perfect as I would have liked, but I was told that EGFR (the trouble molecule) can be pretty difficult to obtain good bands for, even by experienced lab workers. Part of the problem is that it is relatively low in abundance in comparison to pERK, and also, as a membrane protein it is very difficult to extract and analyse accurately. Usually, you require much more advanced techniques to access membrane proteins, but it didn’t warrant the effort for such a short project. As the main goal is to analyse the behaviour of ERK itself, it is not too much of a problem.
I have begun writing my project up now too. So far I’m pretty proud of it. I feel like I have done quite a good job and so I hope that the UTRIP organisers and my professors feel the same! I am considering uploading a copy of it to here when I’m done but I’m not really sure whether it’s allowed. Hmmm.
Outside of the labs this week I haven’t really done much. I’ve worked rather late each evening to try to get everything finished, and when I’ve arrived home I’ve been too tired to do much else. I’ve started watching “Breaking Bad” on the internet, which is absolutely amazing (and I completely recommend it to anyone reading this… which is odd because I genuinely don’t watch a vast amount of telly – I’m more of a reader!), and then have just been going to bed.
I finished reading “Paper Towns” by John Green – I can’t remember if I mentioned that before. It was amazing though and I’m a little sad that I’ve finished.
Oh… and I’ve been feeling quite sick for the past 2 days. I woke up this morning and yesterday with a fever and cold sweats, even though the air conditioning was on full-blast. Not too fun. Hopefully I’ll feel better soon.
Last night, a lot of UTRIPpers went out to celebrate finishing their programs. Literally everybody except for me went by the sounds of it. I’m really sad actually because I’ll never see some of these people again! Why didn’t I go? Well… I was working on my project. Yep – I’m a good student. Yep – I’m a bad friend, evidently. I wish I’d gone. I regret it actually. Everybody came up to my room to come and get me but I was sat in my pyjamas writing my report and really felt like I had to work. My presentation is on Tuesday and I’ve done so little to prepare because I hadn’t even got all my results back until today!
In my defence, I didn’t realise that so many people had finished and were leaving. Had I known, I would have put my project on hold for the evening and gone for a quick drink or a bite to eat.
So… I guess that’s everything. This was pretty short for 4 days but all I really have to say is: I spent a long time in the lab, my first practical yielded uninformative results; my second practical yielded slightly better results; “Breaking Bad” is awesome, and I’ve done little outside of working.
Over and out!
pseudopsience 