User:Linh N Le/Notebook/2009/06/11: Difference between revisions
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*Currently reformatting the HUGE drives, 1hr in and its at 16% | *Currently reformatting the HUGE drives, 1hr in and its at 16% | ||
*I hope the RAID 1 system is easy to get back in order | *I hope the RAID 1 system is easy to get back in order | ||
*time: 4:12pm, 67% done with reformat | |||
**Still have to install windows :( | |||
*computer done installing at 5pm | |||
*nothing comes up when i log in as the admin | |||
**The problem i think is that the install disc that koch made does not have all the drivers that this computer needs | |||
***I think this b/c although it is hooked up to the internet, i get no connection or response from the ethernet cable. | |||
*Will try the drivers cd 2morrow or dl a fresh batch and send them over via usb stick | |||
==Acetic Acid== | |||
*while waiting on the computer, i will help brigette make MT's | |||
*she is mixing up the acetic acid | |||
*Note ok MT's: I will be using the BRB80T that Igor and I made earlier this week (monday?) | |||
**it has been in the fridge all this time, so i think its still good | |||
**mixed it up using the vortexer (koch mentioned that taxol can come out of soln if it sets) and am leaving it on the counter for ~20mins to warm up before adding it to the MT's | |||
*Eventually i will remember the recipe for making MT's but for now I had to look it up | |||
**As of this monday, its 2ul of tubulin and 198ul of BRB80T (premixed) | |||
*Tubulin + brb80t mixed up | |||
**Andy is messing with the microscope right now...he has the translator stage off, so I wont bother trying to look at the MT's until he gets back and gives me the ok | |||
*looked at the MT's, no antifade. They are there. They fade in ~.25s | |||
*Now to add acetic acid to the MT's | |||
**Brigette made .273M Acetic Acid soln (w/DI H20) | |||
**She wants to add 3ul of this soln to the ~200ul of tublules (technically only 195 since i took 5ul to look under the scope) | |||
**Scratch that. We made a soln of 100MT to 3ul of acetic and 100ul MT to 1.5ul acetic | |||
*Right now, brigette is looking at the 3ul aa soln | |||
**They are still photobleaching quite fast, so it is really hard to tell if the tubes are breaking apart or not | |||
*Now trying it with the stock of AF that brigette made last week | |||
*Doing | |||
#10ul of the tubes + 3ul acetic acid | |||
#2.5ul of AF | |||
#1ul of 2M dextrose | |||
#80ul of BRB80T | |||
*took the soln above and gave briggete and sample to look at | |||
*took the soln above, added 3ul more aa to the whole stock, make sample for brigette to look at | |||
*note: I only used maybe 1/10 of the AF cocktail today | |||
**refroze the aliquot, to save money, marked the aliquot with "thawed 6/11" | |||
**I hope AF is ok with being thawed, refrozen and thawed again | |||
*Brigette and I have spent the good part of an hour trying to image our new samples. | |||
**~45mins after creation, I took a look a new sample of the AF+MT(3ulaa)+dex+brb80t+2ulaa mix and saw some floaties. | |||
**The AF didnt seem to work very well. The background went black too fast! | |||
*Getting frustrated at the fact we could not see anything, I went ahead and sampled out the pure tubes +1.5ul aa | |||
**We saw plenty of tubes, and even though they do fade quite rapidly, you can still see them faintly in the background | |||
*Conclusion of the day: AF does not screw up MT. AA does not screw up MT. Mixing the 2 messes everything up! | |||
*In a last ditch effort, Brigette added 10 more ul of MT(with 3ul of AA) to the AF mixture | |||
**Hopefully more MT's allows us to see some | |||
*On a side note: we really need another microscope as Andy came back into the lab and wants to work on the tweezers, but we are still looking at samples! | |||
*MT's, no antifade, with aa, can see tubes. MTs with AF, with aa, still fades badly, but not tubes. "bad" AF that doesnt stop fading, +MTs, can see tubes, but they fade fast (as we saw last week). MTs + aa +AF (whether or not its good b/c it was in the freezer, or even if I messed it up by not thawing it right) no tubes. aa+af=bad for tubes. | |||
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Revision as of 18:41, 11 June 2009
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