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.