User:Linh N Le/Notebook/2009/08/28

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*added 25.8mL of 1M NaoH to get it to a Ph of 6.89
*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  
**Thats 12.9mM of NaoH in 200ml soln of PEM  
 +
==DI H<sub>2</sub>O==
 +
*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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Revision as of 14:30, 28 August 2009

Project name Main project page
Previous entry      

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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