9:30a – 4:00p

Computer Management

attempting to give users rights to share their hard-drives on the network without having full Admin capabilities to do anything they want.
Tried adding my ‘User’ account to these groups
- “Performance Monitor Users” — can access performance counter data locally and remotely.
  - This doesn’t actually let me launch task manager without Admin rights. Not too surprising, since I can kill tasks from task manager.
  - Can’t even access the windows program “Performance Monitor” without Admin rights.
  - Can’t launch the “Resource Monitor” without admin rights either
- “Access Control Assistance Operators” — windows / google seem to provide no information on what this really does.
Can now open preferences, still get asked for admin password to try to change advance sharing file permissions

Chromatin Project

have staining now of C01 in both new cells and old cells
- looks better in new cells
- neither is comporable to earlier stains or restained c8 cells.
- some cells have strong/decent stains and some cells lack staining.
- pretty sure there’s higher background, different quality. May relate to lack of RNase treatment in new cell round. —

should have checked — C01 is 240 kb Y. We probably do have mRNAs in cytoplasm.
don’t have spots in every cell, when we do have spots the cytoplasm is dimmer. maybe I’m just staining RNA. Can treat these cells and see if we lose stain?
previous C01-C08 batch of cells were first imaged 1 day after I made new RNase solution, so I don’t have it for sure in my notes but I’m pretty confident these guys didn’t skip the RNase step. So current stains are consistent with not denaturing the DNA, except that a side by side re-treatment of C08 cells does denature for strong incorporation. So something is still different in the cell prep that affects the denaturation, rather than the denaturation itself.
maybe RNA also dilutes out concentration of local probe (I kinda doubt it, probe concentration is pretty high since whole hybe soln is visibly blue).
maybe somehow cells are over-fixed and won’t denature? I think I’m very consistent though on fixative and fixation times…
I guess I often let them sit longer in the glycerol than the 30 min I have been doing — I sort treated that as a minimum to get properly equillibrated rather than an exact time step.

RNase treat current stained C01 new cells, see if foci are lost. Treatment started 1:20 pm.
RNase treating new cells prepped with new buffers. Will then do extended pre-hybe.
95C denature of RNase treated cells prepped yesterday and restain with C01.