To Do
- Write my last official notebook entry as a KochLab member :(
- We are running out of PEM, so I would like to make some more before I go
- We got motility working yesterday, so Andy may want to continue to pursue that
Inventory
- Brian had to leave, but i did give him a crash course on how to use the inventory page
- Most of the pages are self-explanatory
- Also note that chemicals and software inventory require an update to the google docs page
- The optics section of the inventory is blank
- My idea was to have it as a static page that just listed important opto-mechanics
- Perhaps we can have a check in-check out system so we can keep track of who took what where and when
PEM
- Andy, Brigette and I made 205ml of PEM a month or 2 ago and its getting low
- I will try to make 200ml more and follow the exact steps I took last time
- I will need NaOH and possibly HCL to titrate the buffer
- We should have plenty of normal soln left from earlier buffer makings
- We have 4 tubes of NaOH but no HCL, lets hope I dont screw up the titration
Procedure
- Following what I did last time, I want the following:
- 1mM MgCl2
- 1mM EGTA
- 80mM PIPES
- I am using the following chemicals:
- 1M MgCl2 in 50mL falcon tube made 2 years ago ( we used it in the motility assay yesterday and it was fine)
- For 1mM in 200ml, I want .2ml (200uL)
- EGTA powder, working weight 380.35
- For 1mM in 200ml i need .07607g
- PIPES powder, working weight 302.37
- For 80mM in 200ml, I need 4.83792g
- I will wash out a 250ml glass bottle with DI water then use DI water to suspend the salts
- I will use the autoclaved DI water I made a month ago, that way we use it a up a bit since we have a fancy schmancy DI water machine
- I filled up the jar with ~150ml of DI h20, now to add the powders
- After addition of powders and washing it down with more DI water, I'm about 175ml
- Powder Concentrations used:
- 200ul MgCl2
- .0768g EGTA
- 4.8387g PIPES
- Note: I used the "DI Water" in the spray bottle. It may be tap water, Andy and I both dont know who filled it up with what. Last time I made buffers I used the water in the spray bottle, hopefully it still works
- Now to titrate
Titration
- Starting Ph
- added 25.8mL of 1M NaoH to get it to a Ph of 6.89
- Thats 12.9mM of NaoH in 200ml soln of PEM
DI H2O
- I went ahead and emptied out all the squeeze bottles that are labeled DI water and replaced it with the stuff from our DI water machine
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