I'm firmly establishing my own well-worn ass-groove in the leather chair that goes with my desk in the cholera lab. One thing that annoys me about my desk set-up is that the chair is fairly wide and it has armrests. The armrests are so wide that every time I scoot in toward my desk, one armrest or the other hits the side of the desk. Every. Single. Time. I've got half a mind to take a damn screwdriver to school and remove the fucking armrests. If it's annoying me now, imagine how I'll feel after five years of it. Maybe I'll wait until the PI has agreed to accept me as a full-time lab member and gives me a key to the lab before I start disassembling his furniture. Since I didn't put in as many hours during my rotation as I should have (to avoid failing my classes, not making it to prelims, and subsequently being kicked out of the program), the PI is letting me work in the lab over the summer on a "trial basis" to make sure that I will actually show up to work and generate some decent data. So I'm technically not a full member of the lab. However, I have no intention of being kicked out, asked to leave, or whatever. In my mind, I'm a permanent lab member - it's just not official yet. Unfortunately, I'm reminded of my provisional status whenever the PI sends an email to the lab, but only carbon copies me on it. I don't really understand why that arbitrary distinction has to be made in stupid two-sentence emails stating that we're having lab meeting this week at 2:00 pm - like we do just about every week. That bothers me. Alright, that's enough of the complaint department - I'm starting to feel really pissed off about my stupid provisional status, which will get me nowhere fast.
I had a good productive day in the lab today. It almost felt like it did when I worked in Jim's lab at Eastern, when the science determined what time I went home for the day- not sheer boredom. My PI is off this week, so I don't have to worry about him keeping a surreptitious eye on me. Yay! I feel like I made a butt load of media, but I don't think I actually did. Let's see, I made a liter of 5X M9 salts, 100 ml of 10X PBS, a couple of 90 ml dilution blanks, 100 ml of 20% (w/v) glucose, 100 ml of 1M MgSO4, and finally a liter of complete M9 minimal medium. The shelves atop my bench are becoming beautifully lined with bottles of stuff. Most of the bottles are identical, each containing exactly 100 ml of whatever potion I whipped up - just like everyone else's benchtop shelves. I'm telling you - I really feel like I've joined a cult. Thankfully, no one is sporting any forehead swastika tattoos and wild eyes or wearing all black outfits with gleaming new white Nike shoes. All the perfectly filled and arranged bottles make me feel like I'm doing real science. M9 is a royal pain in the ass. I think that alone took up half my day - just autoclaving and cooling to room temperature. Fucking ridiculous. But, I had to make it because that's what I'm growing my WT cholera and ToxT deletion mutant cholera in overnight for tomorrow's zebrafish infections. I'm just trying out a very basic experiment to see if it works and to see how long the whole ordeal takes. I'll have two groups of four fish each in separate beakers of 200 ml sterile saltwater. I'm going to add my overnight cultures: WT (cholera toxin-producing) to group 1 and ToxT mutant (no cholera toxin production) to group 2 and incubate the fish at 28 degrees for 24 hours. Then, I'll euthanize the fish and collect the beaker water. The plan is to concentrate whatever protein is in the 200 ml of fish water by trichloroacetic acid precipitation - which I've never done before, so will probably screw it up. The multitude of proteins precipitated from the fish water will be used in a cholera toxin ELISA, which I will also likely screw up in some way. The idea is to see if cholera toxin is present in the fish water. In previous experiments, when naive fish were placed into water conditioned by other cholera-infected fish, the naive fish died. It was assumed that the naive fish were killed by cholera toxin that had been secreted into the water, but no assays were done to confirm this.
Oh, shit. It's almost 12:30 and I haven't had my nightly Ben and Jerry's Coffee Toffee Heath Bar Crunch ice cream gorge session yet. And I haven't finished the paper I'm supposed to read for my virology discussion tomorrow during lunch. And I'm still Jonesing for more of my second read of "Fifty Shades Darker" before I fall asleep. I just love trashy sex and romance novels (with a smattering of BDSM thrown in, as the case may be) - I am certainly NOT a fine literature critic, but I like what I like and, embarrassing as it may be, that's good enough for me. And Chris :)
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