Would you like a bag for your tampons?

Ok, now that I've made all male readers squeamish, I'm going to tell a story. I was at the grocery store this morning buying, yes, tampons. There was only one cashier, a middle-aged rough-looking guy. I got in line, and immediately behind me two firemen got in line with their cart full of groceries for the firehouse. Embarrassing enough already, sure. I put the box on the belt, no other purchases, and the cashier scanned it. "Wow," he exclaimed, "did you see the price on those?"...he motioned to the LED screen. It read, "$1.20." Now, the sign on the shelf did say "$4.49" but it's not like I care. This isn't exactly the type of item you shop around for the best price on. "Um, that's um, pretty good," I say. "They must be on final clearance or something, you lucked out!" he exclaims. "Um, yea, I guess so," I reply. Now really, do we have to stand here having this conversation on what a great deal I'm getting on my tampons?...the whole time with these two firemen unloading their cart on the belt right next to me? After I handed him some cash and he handed me a receipt, he finished by asking, "would you like a bag for those?" No mister cashier, just duct tape the box of tampons to my forehead with the receipt attached so I can run into the parking lot and let everyone else know what a great bargain I just got on tampons!

Elevator Master

I just joined a gym for the winter. This is one of the luxuries that the ARCS scholarship affords me, quite literally. I went there for the first time after work yesterday. All of the cardio equipment is on the 2nd floor in a loft-ish area overlooking the rest of the gym. As I jogged along on the treadmill I watched as 4 girls came up the elevator...and got immediately on stair climbers. ?!?!

Spoke too soon

Yup, I spoke too soon. Got too excited. I must always remember...PCR-SSCP is cursed in the hands of researchers in lab number 7309. Cursed. The image looked great. The overstaining problem appeared to have been solved. I changed the resolution setting, and clicked go for the high resolution image of my gel to be scanned. This is what I got:



Waaay too dark! Not enough contrast to run and decent image analysis and develop electroferrograms. And this happened repeatedly for the next 4 hours. Garbage. As a last ditch effort, I turned the scanner off and repositioned the gel in a different corner of the scanner:

Much lighter background....but my bands were lighter too...disappearing. It had been 5 hours since I unhooked the voltage on the gel, and now the DNA was starting to diffuse through the gel. These faint fuzzy bands are worthless to analyze. I tried restaining to see if I could get the bands any darker. No good. The stain wasn't faded, the bands were just diffusing and fuzzing out. Boo. PCR-SSCP strike 3. This is the closest I've come to getting actual results. The bands I see confirm my hypothesis, so I know I'll be happy with the result and that it fits in nicely with the rest of my data. But this is the one last crowning experiment that I'm trying to finish so that I can include it in a paper. And I really want to get the paper submitted before I start taking my prequalifying exams this fall. To top it off, I'm just about out of the DNA I was using in these experiments. That means I have to go back to square one in order to repeat the SSCP. It'll take at least a week. Grr. That's the last time I celebrate until the data is actually in hand.

Success!

Finally!!! This image looks better to me right now than da Vinci's Mona Lisa, or Van Gogh's Starry Night. It's a picture of a PCR-SSCP polyacrylamide gel that I ran this week. This is my third attempt at PCR-SSCP, and if this didn't work out, I was going to have an aneurysm. It takes days to prepare everything needed for this experiment, extracting the DNA, running the PCR, converting to single-stranded DNA, pouring the gel, running the DNA on the gel, and staining the gel. It's the last part that has been hindering me for a few weeks now. It should be the easy part, but it hasn't been. My entire gel was turning black, not just the bands of DNA. No one in the department could figure it out. I stumped tech support from the stain's maker. My advisor said to me, "Chris, I encourage you to come up with creative solutions, not creative problems." Sooooo frustrating. Turns out it's alot of little things....like that the stain that is designed to bind "exclusively" to DNA happens to also bind to the type of plastic we put the gels on in order to scan them. Once bound, it fluoresces...and appears black in the image. If the gel isn't completely washed of the stain before going on the plastic, poof! A big black mess. Keeping the stain off the plastic just about takes more effort than the entire PCR-SSCP process before that. Lots of other little things factor in too. The staining process is definitely much more art than science. None of the manufacturer's suggested staining times or concentrations came even close to working. This here is just the prelim, 2 minute scan image. A much more detailed and resolved 30 minute scan is going on right now. This has been such a headache. I think I deserve a beer! Maybe two.