User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/09/14/Getting motility to work again
To do
To keep moving forward, I'm going to keep writing my "To do" list at the top of each notebook.
- Hydrophobic surface.
- Hydrophobic surface with a groove.
- Corn starch.
- Potato starch.
- Gluten/flour.
- Silica nanoparticles.
- Silanized surface. I'm not sure how to do this one but apparently lots of people do it to attache DNA to a coverslip.
- Lipids with PEG on a hydrophobic surface spin coated.
- Better spin coating of lipids on coverslips.
- PE lipids?
- Gelatin?
New stuff
Okay, Koch told me to try new kinesin today with the stuff I had made with the possibly questionable buffer or Taxol solutions. When I went to look at the stuff, I found that there were no microtubules in solution.
Rather than try and figure out what is going on, I have decided to make everything new. I did find out that DMSO, the stuff used to reconstitute Taxol, could have gone bad. The MSDS states that DMSO is very hygroscopic and should be stored in a cool dry place, preferably under nitrogen. Well, our DMSO is in the -80˚C freezer and has frost all over it. While this may not be a problem in the freezer, it is a problem when taken out of the freezer. I am no longer sure about the quality of our DMSO and thus I am not sure if our Taxol solutions are still usable.
The major reason why I believe the Taxol solutions are not working is because of the fact that I polymerized microtubules on Sept. 08, 2009. They should still be viable and yet they are not. SJK 23:10, 14 September 2009 (EDT)
actually, that's not surprising at all. We never used MTs for more than a day unless we were specifically studying their long-term stability. Taxol is good, but not that good (it's not covalent). Could this be your problem???? I would never have expected your motility assays to work today with MTs that were almost one week old!
I have decided to get some DMSO from CRLS. It comes packaged in 5X5 mL ampules so they should be good forever if they are not opened. I also got some vials to put the stuff in so it can be dessicated and stored in the -20˚C freezer.
