User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/09/30/Preparing for experiments

Hmmm

After thinking about it, I am going to make a page that describes what I am doing in the KochLab protocols section. That way I won't have to write what I am doing in this notebook page and there.

Tweezers

I'm also helping Ant with the tweezers.

ATP

Continuing with my stock preparation, I have made several aliquots of 100 mM ATP in PEM. I just weighed out the appropriate amount of ATP and put it in PEM. ATP auto-hydrolyzes and the amount of hydrolysis is pH dependent with less hydrolysis occurring at higher pH. Books will tell you to store ATP in Tris at a pH of 8 but I found a paper that shows storing ATP in a buffer with a pH of 7, is not that different. Plus, I'm sure that people say to store ATP in a pH of 8 because they don't freeze it. Not sure but if it starts to go south then we will know in the future to store it in Tris.

I made an initial stock solution and then aliquoted it into 10 µL aliquots. To lessen the amount of time the ATP was at room temperature, I used the freezer box to store my stock in while I aliquoted it into tubes and flash froze them in some LN2. They are labeled and now in the freezer storage box shown above.

GTP

GTP is used to polymerize tubulin. It is needed also to store the tubulin in "ready to make" aliquots of tubulin. I believe that it was decided that we should store GTP in PEM with an additional 1 mM of MgCl₂.