Wednesday, October 04, 2006

The 12hr Time-Point

Its weird—what Im doing right now in the lab is exactly what I was doing this time last year. But this time last year I was a retard that didn’t know how to pour an 0.8% agarose gel. Seriously, my boss had to show me how to do that (and he was very sweet about it, btw. Didn’t even laugh when I tried to ‘write down the protocol.’ **blush**). Now Im doing these insane experiments by myself.

Including the dreaded 12hr time-point. Ugh. No matter how early you start this protocol, the 12hr point is always in the middle of the night!! ARRRG! I go to sleep 30 minutes ago!

So Im typing this blog entry to keep me awake until I have to drive to work and blow stuff up for 45 minutes. Yes, blow stuff up. Blow radioactive stuff up. I said these experiments are insane!

I infected some cells with a virus a couple days ago. Let the viruses get real comfy—let them set up shop, getting the cell to make its proteins for it. Hehehe But this morning, I took away all the cells’ food. I starved those bastards so they couldn’t make any proteins. Hehehe Then I gave them food, laced with radioactive Methionine and Cysteine amino acids!! Methionine and Cysteine are the only amino acids that contain sulfur, so we just exchanged a regular sulfur atom with a radioactive S35 atom—that way we could find out exactly what proteins the cells were making, because all the proteins they make will be radioactive! And those cells are making viral proteins :) We wanna know how much of a certain viral protein is getting made, and how fast its getting chopped into two pieces that the virus really needs to stay infectious.

Well we can collect all the proteins the cells make by blowing them up and collecting their innards (my favorite part, personally hehe). We blow them up right after we give them radioactive food, an identical batch of cells 2 hours later, another one 4 hours later, etc etc etc up to 24 hours. Again, we want to know how fast this protein were interested in gets chopped into two, so we take a snap-shot every couple of hours.

We can pull out the viral proteins we are interested in by making them stick to beads with special antibodies. These proteins get run out in a gel to separate them out by size—put a piece of film on the gel, let it set for a while, and the S35 excites the film when exposed to light, and you get a sweet ass gel that you can get lots of AWESOME data from!

… But that will be like, a few weeks from now. Sorry. :(

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