So this week has been the start of my placement. As I already know my way round the lab (I'm am so grateful for this, it usually takes weeks) I have been able to get straight on with doing things! I've been growing up cells, mainly E. coli mutants, and adding proteins to them. My agar even worked. I have a history of agar hating me. I can make it fine, and if it has come straight from autoclave it's great, but that rarely happens, so the 'melt in the microwave' technique is the best option, and either I don't heat it enough and it's lumpy, or I heat it too much, then panic that I'll denature the antibiotic and then leave it to get too cold, and it's still lumpy. However, thanks to being made to practice earlier this year, which I'm very glad about, this week's agar worked really well.
This brings me on to word of the week, which is Kanamycin. I know it's only an antibiotic, but it sounds great!!!
Panicked over my SDS page gel yesterday, which unusually decided it was going to take twice as long as usual to set. The gels ran really well today (well after I was reminded to boil the samples anyway). For anyone who doesn't know what the gel is, it allows you to separate proteins of different molecular weights using an electrical current. The smallest proteins travel faster so end up at the bottom. You can stain the gel and compare the bands that appear with a marker that has known molecular masses in it. This helps you to identify if the protein you're looking for is there or not.
Today I was particularly happy as I started my first western blot ever. This is where after running proteins on an SDS page gel, you then use electricity to drag the proteins onto a sheet, to which you can add antibodies to probe for the protein you want. I was surprised to find that the process involves milk powder!
Feeling a bit heady due to pouring over my gel to see if we could find the protein, and breathing in a bit too many gel destain vapours!
I've also done loads of reading, and feel a bit more clued up on the subject of my project, there are so many things to know about just one protein, you'd be surprised.
I included teenagers in the title, as today we had a school visit, which the whole lab helped out in. It was a microbiology based set of practicals. I took great delight in telling them they were looking at the plague, anthrax, MRSA, TB, and various other lovely diseases. Not so sure they appreciated it so much... Also got the opportunity to judge the 'design a microorganism' competition along with Alex, a masters student in the lab. There were some amazing ones, including ones that give you a six pac; clean your intestines; turn your hair into cheese; made you happy; and made your eyeballs explode, but most of all, made you die a slow and painful death. We ended up choosing the two that didn't actually kill you. Had a really fun day and managed to do quite a lot of lab work in between.
Looking forward to finishing my western blot tomorrow. I will find out if my first set of experiments have worked. So far the only thing I really know is that my basic lab skills haven't been terrible so far! I'll write another post, hopefully early next week about how it's all going, and perhaps make it a little shorter!
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