Isn't Grad School FUN!!!!

Let me attempt to sell you on Grad School by describing my Tuesday:

First off, I finally roll in around 9:30 which is later than I wanted to be there, but if I'm home when Laura comes back from her bike ride I am compelled to wait for her to shower and eat second breakfast before I drive in. To do otherwise would be mean. Usually I'm out the door but because I was still recovering from my marathon driving weekend I was still in my pajamas at 8:45 or whenever she got in. Oops. By the time I got to work DLC had already killed 3 fish and hadn't saved me any livers. Humph. That's okay, I'm reworking my protocols anyway. After nearly a year at the same grind you'd think that I'd have this enzyme assay thing all wrapped up by now, but no. Grad School time is 1,000 times slower than real world time. Or maybe it's just me.

So I made some new reagents. Because if I know one thing about molecular biology it's this: When stuff isn't working, your stuff is probably contaminated/yucky/sabotaging your experiments. So, I made new buffer, new poisons (just in case THOSE go bad) and new water (we make that too). Then, I pulled a Complex IV assay kit out of the freezer and prepped that for a go at testing whether the mitochondria we're preparing are really even intact. Turns out they are and running an enzyme assay that worked was a real morale booster even if it wasn't for the enzyme I love to hate: Complex I. I was buoyed by my success and decided to have a go at Complex I quantification. I used some of the tricks suggested by the Complex IV assay kit. I mixed the master mix at room temperature, I initiated my reaction with NADH and I used the same protein concentration that I used for the Complex IV assay. It was a disaster. The absorbency data were all over the place, it looked more like a jack-o-lantern's mouth than a nice even, linear, slope.

At this point, when I've spent all day of spinning and pipetting and thawing and homogenizing and freezing and standing on my head while spitting nickels I throw up my hands and say IT DIDN'T WORK! And I vow to try again, might work next time. Like, maybe, tomorrow 'cause damnit if I'm not DONE for today. Yesterday was different, I had the sweet taste of technical replicates with no variation and huge differences between my experimental and control slopes. I had it goin' ON! I could do it! So, I tried again with 10X protein concentration and would you know it? IT WORKED! For the first time EVER my slopes looked like what I expected them to.

Ahhh, I breathed a big sigh of relief and rushed off to a meeting. I was driving back to the lab to get some more assays under my belt when Laura called. "Have you seen your email lately?"

"Uhh, no I've been out of the office" She knew this...

"You're giving your student seminar on Friday" She had heard through the grapevine already.

I can never catch a break. Instead of celebrating my assay success and repeating and repeating and repeating while dancing and rubbing some good juju on my spectrophotometer to keep the good times rolling, I'll be chained to my desk trying to recreate some crummy talk I started working on in November. What am I supposed to say? Why is this important, I forgot a little......