another life blog

Drosophila melanogaster leg imaginal discs like to “stick” to different surfaces ~ especially little plastic replacable pipet tips. To prevent the discs from sticking, we add bovine serum albumin to the different solutions. This special beefy protein, would charge the walls of the different surfaces, and repel the discs from sticking to the surface — wonderful, isn’t it ?

However, to prepare my sample for high performance liquid chromatography, I must wash the imaginal discs from the incubation medium with PBS, Ringer’s, or distilled water. Now, if I wash the imaginal discs in the 1.5 mL centrifuge tube, I would need to centrifuge in between washes – and centrifuging would break the imaginal discs. So, I must wash the discs in the culturing well. If I wash the discs in the culturing well, that means BSA would no longer be present in the well, nor in the pipet tip to prevent the imaginal discs from “sticking.” So naturally, I thought — let’s add BSA into the wash!

Wrong. BSA would disrupt the HPLC result because we’re getting a ratio between amount of drug versus protein weight. BSA is a protein, thus we can’t have BSA in the wash. So, what’s a man to do? Hope I have enough imaginal discs that some would be suspended in liquid when I eject them from the tip into the tube.

This is not the worst part. The worst part is after sonication and the final centrifuging, there’s no pellet. No pellet of protein at all! So basically, I can’t get a ratio, and I wasted 5 hours for… naught. And .. I’ll need to do it again. Yup, it’s somewhat frustrating…

This entry was posted on Friday, March 31st, 2006 at 2:39 pm and is filed under Undergrad @ UCF. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.