User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/11/09/Water isotope study
50% HPEM 1st try
I was able to find MTs within 60s from the bench to the microscope. It's interesting because these guys are pretty much stationary when I first inspected them. They are moving albeit rather slowly. I'm sure as the temperature increases they won't be so slow which also means that they won't break apart.
Yep, the 2nd ROI has no problems what so ever with it.
This may prove to be interesting. I don't think that I prepared my PEM-alphaA properly. I think I just made some PEM with 1 mM ATP in it. The next 50% does have proper PEM-alphaA.
50% HPEM 2nd try
So I made some more PEM-alphaA and stuck my kinesin in it. We shall see if there is any differences in speed between the two assays.
This is very interesting. This assay didn't work. I'm not sure if I put kinesin in it then. I guess I'll try it again and see what happens.
Ha! I'm an idiot. I didn't add kinesin to the slide. It's there now and everything seems to be working just fine.
In movie 10 there is a MT that is well over 100 μm long.
70% HPEM 1st try
No problems initially. And none at all. There was no breakups here.
I am a bit concerned about the number of MTs that are moving and their minus ends are coming off of the kinesin surface. I'm not sure but I think the kinesin may have gone bad. I'm going to switch to a new batch of kinesin and see if the same thing occurs.
70% HPEM 2nd try
So far so good. They are moving slowly in the 1st ROI as expected. And, they are breaking.
The rest of the assay looked great and I got much less minus ends in solution.
80% HPEM 1st try
Zero problems. I wonder if the higher concentration of D2O is helping prevent the initial breaking I have been witnessing in the assays.
80% HPEM 2nd try
Everything seems to be moving smoothly. Except for the 1st ROI of course.
20% HPEM 2nd try
Well, this assay failed miserably. I'm not sure why it did but I'll have to figure it out. I'm going to try another assay with the same stuff to see if indeed there is something wrong with the sauce.
Nah. I'm quiting experiments for the day.