User:Linh N Le/Notebook/2009/06/16

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LN2 Container

  • Working with LN2 yesterday to flash freeze the tubulin made us realize that a container to transport it into the lab would be much more useful
    • Very apparent when trying to haul it in with the flat, shallow, red ice bucket and having it almost spill everywhere
  • For $3-500, it did not seem like a worthy investment
  • Instead, today, we will take the black ice buckets and fill them with LN2, then transfer it in the lab as needed
    • useful the the spigot is only ~2m away from the back door
      • Possible to get a feed inside the lab? (just a thought)
        • Steve Koch 23:54, 16 June 2009 (EDT): Yes, that may be possible. Definitely $300 is worth it if safety is an issue. As I see it now, a couple trips with small bucket seems reasonable and safe. But let me know if not!

Videos from yesterday

  • I am moving them to the webpub as koch advised me to.
  • Moving to \\controller\webpub\files\movies\MT movies
  • Should be accessible via kochlab.org

Video Card

  • The subject of videos reminds me that I should go and make sure Tamra gets my order in
  • I guess i'm really good at catching her when she's not around
    • will send an email instead

KochLab Inventory

  • Koch wants me to go over the inventory page and see what else needs to be inventoried
  • something we may want to add as inventory are:
    • Wetlab shelves
    • The fridge (VWR next to -80c freezer)
    • Fume hood
  • I guess that means I will need to make new templates to go with the new inventory areas

Wetlab shelves

  • I want to me a naming convention for these shelves
    • Looking at the shelves with your back to the main sink towards the fume hood
    • Upper left 1, below that 2, upper right 3, below that 4
    • please also mention if it is in front or back
      • i.e ph7 buffer Wetlab shelf 2 front

New Templates

  • I made 3 new templates for the inventory page
  • Mark a page with the following templates to indicate where it is
  • example marking {{KL 80|b=shelf 2}}
    • Reorders {{Reorder}}{{KL Inv}}
    • -20c {{KL 20}}
    • -80c {{KL 80}}. Also include |b=black or |b=shelf 2 for location
    • VWR fridge {{KL VWR}}. Also include |s=left or |s=right to indicate the side of the fridge it is in
    • Wet Lab {{KL WL}}. Also include |s=1 or |shelf=2 for shelf spot (see convention above)
    • Fume Hood {{KL FH}}

Kinesin + tubes

  • we polymerized another aliquot of the RMTs
  • Koch and brigette made a new stock of BRB80T
  • I tested the tubes with an old AF we used yesterday and refroze
    • from what we can tell, it works ok, but not as good
      • fades faster + there are broken tubes floating around
    • turns out this AF was the one i used when we first tried AA.
      • Brigette thinks that it is bad AF to start with, so we dont know
  • thawing a new AF to test this theory
  • koch has also made the new computer the camera computer, so we are playing with that now
    • Works as intended

kinesin

  • Koch and brigette worked out the formula for the motility assay
  • I dont have the exact details, they are in Koch's notebook (i will link later) (User:Steven J. Koch/Notebook/Kochlab/2009/06/16/Motility assays)
  • They made the casein solution (From the powdered milk) and the motility soln
  • The kinesin that Koch used was some old stuff from ~5years ago
  • We made a dual chamber slide
  • One had casein + motility soln (tubes, af, atp, etc) + kinesin
  • The other only has motility soln (i cannot remember if both had casein in it)
    • The control (tubes only) showed us tubes that were stuck mostly to the glass
      • on that note, i dont think the control had casein
    • The kinesin side had alot more loose tubes and alot more tubes that were only stuck at one end and flopped around.
    • The ones that were stuck to the glass we watched. It did not seem that they were moving at all
  • Trying again, we made another dual chamber (note: makes sure the channels run along the length of the slide, it makes things much easier, especially when looking with the microscope.
    • One chamber had the original kinesin mixture as a control
    • The second chamber had ~10x more kinesin
      • I personally only got to look at the 10x sample, but brigette says the 1x looks exactly the same
      • The 10x seemed to have much more "stuck" tubes (That is not moving around and completely stuck to the glass)
      • Other than that, i saw no movement
  • Some thoughts
    • Is the casein coating all the glass? I know that tubes dont like casein, but maybe they are sticking on areas that dont have it coated on there
    • Will kinesin bind tubes, even if the kinesin wont walk? My thoughts are no, but the idea came up with the 10x concentration. Perhaps the kinesin are in the casein and are acting like velcro, just sticking MT's down
    • No huge surprise that 5yo kinesin stored at -20c (instead of -70c) doesnt work too well
  • Movies were made, I will upload these to the webpub with the ones we made yesterday!
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