Monday, June 21, 2010

What the heck do I do at my job anyway?

I've worked at Karmanos Cancer Institute for almost nine months now as a research assistant in one of the breast cancer research labs. I got the job in a roundabout way that involved my step-aunt's best friend, the CFO of Karmanos, passing my resume to my now-boss who called me because I was apparently well-qualified. The upshot being that I work as a research assistant now, which means I do most of the work without getting much of the glory. Oh well. It's not forever.
At the moment, I am in the midst of a rather large experiment involving a panel of genes that are candidates as either oncogenes or genes that are regulated by an oncogene. What I do with these candidate genes is make an shRNA against each one and infect my target cells with said shRNA to knock down the gene's expression. Then I wait and measure growth and viability with a luciferase assay to determine the effect of knocking down the gene. In this experiment I have 20 genes and 148 shRNA constructs. That means I spent all of last week making 148 viruses to infect my target cells with. It was unpleasant. This week I get to infect my target cells, which are MCF10A and SUM225 lines. I do the infection in triplicate, which worked out to 444 wells, or four and a half 96-well plates. It's also unpleasant, mostly because I have to add the virus one at a time and can't use a multichannel pipettor. After that, I wait and see what happens. Our constructs have a GFP tag in them so after 24-48 hours I should be able to see some of my infected cells glowing green, and that's how I know that the infection worked.
Anyway, that's what I'm doing at the moment. Previously I did some work on PHGDH knockdown and insulin independence, which was mostly growth assays, Western Blots, and qPCR. I'm not fond of Western Blots. I'll complain about them another time. I've actually learned a lot in the nine months I've been working, and I feel a lot more confident that I can do science and could have a career as a research somewhere. Although I feel my leanings are toward some part of microbiology, the time I've served in cancer research will definitely benefit me.

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