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Gels can Smile?
Submitted by Jennifer Newsted on Sat, 01/26/2013 - 11:42am
Bioscience Technology at Portland Community College is getting slippery! As our class forges ahead in the fun filled world of Bioseparations we tackled chromatography and protein gels (protein electrophoresis) this week! We learned how to piece together a gel set-up which is like negotiating a particularly tricky set of Legos. We also made our own resolving and stacking gels! Gels are fun!
There’s a lot going on in running gels. Sure, I’ve seen a picture of a gel in textbooks. Our class even talked about them a bit in Current Topics last term, but I wasn’t remotely aware of all the things that are going on to get that little picture until this week. For instance, the gel filters proteins based on their size. So, in order to get the proteins on an even playing field where their size is the only difference between them, the stacking gel has to denature the protein which causes them to unfold. The stacking gel also changes all of the charges to negative. Once the proteins are treated and stacked, they begin to move through the resolving gel. The rate at which the protein travels through the resolving gel is dependent on the percentage of your resolving gel. The proteins fall down through the gel faster if it’s a lower percentage. The smaller proteins fall down faster because there are less of them to get caught up moving through the gel’s beads. Right, beads. The gel itself contains little tiny beads that we took a look at through the microscope. It made me think of that Price is Right game Plinko. Which, incidentally made me think that my great grandmother would have really loved watching Drew Carey host the show.
So, after playing (and by playing I mean working safely and seriously with crosslinking agents) with some neurotoxins (acrylamide), we had our gel cartridges ready to polymerize so we could move on to loading them. Wow. Loading them. Talk about a counterintuitive procedure. Before you line up your micropipette and pray to the gods of gel comb wells that your eyes do not deceive, you have to pour running buffer all the way up and over the top of those gels you just made. Then you add the loaded tip of the micropipette to the spot you think the well is (ok, I’m exaggerating, you can see the wells, sort of) and carefully load the well.
Our instructor let us go bananas and actually over load the wells so we would run what hopefully would be the ugliest gel we will ever run so she could show us all of the things that go wrong using our examples. I’m still not entirely sure how overloading causes the “smiling.” Anybody?
That was what we did with the end of our week. The truth is, the construction being done at our campus and on our building necessitated the water being turned off. As a result, with no emergency shower or eyewash station, we weren’t able to use any chemicals. Since we still had emergency sprinklers in case of a fire, I asked why we couldn’t just go ahead and work, but light a Bunson Burner and hold it up to a sensor if someone manages to get themselves in a mess. Our instructor knew I was mostly joking. She pointed out that in that case everyone would have to agree to the emergency shower which we just couldn’t get a consensus on. Weird.
While our water was out though we had non-stop lecture/dry demo time. It was refreshing to know that I'm in the right program for me, because I can just about guarantee if it had been all day lectures on anything but biolab fun stuff I probably would have been in physical pain. The lectures were actually fascinating.
We’ve been kind of spoiled in our program by having some dedicated instructors who probably live at the school and are therefore our instructors in multiple classes. It’s a huge bonus to be able to have a professional not only become familiar with your work in the lab, but also task us with the types of questions we will need to ask ourselves as we troubleshoot. We also have had someone we feel comfortable going to when something is just not making any sense.
One of the instructors we’ve been able to become close to is going to be leaving us shortly. And while I am so happy for her to be given what has to be an exceptional opportunity (I mean it would have to be to leave our group! We’re pretty awesome.) I know that am not speaking for just myself when I say that we’re going to miss her and we appreciate her sharing her knowledge (and patience) with us. (Sorry, for being so cryptic but I didn’t get her permission to use her name here. )
The fact is though, that any of the instructors in our program are ready and willing to help us make sure we get the most out of the program and the material. If you're a student at a different school, I'm willing to bet you probably have some of those instructors too. Make sure you take advantage of their willingness to help! Hopefully soon we will be the ones receiving some amazing employment opportunities!
Recipes and protocols from my lab notebook for this week's adventures will be uploaded to my website next week. You will be able to find them at jennifernewsted.wix.com/biosciencetech. Happy learning!