User:Linh N Le/Notebook/2009/07/07

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To Do

  • Camera stuff
  • Kinesin activity assay?

Camera

SJK 23:04, 7 July 2009 (EDT)
23:04, 7 July 2009 (EDT)That's good to know that it works really well wired.  One really important function is the remote camera control: Taking a picture while controlling and viewing from the computer.  Does this work well over the wired?Linh N Le 01:08, 8 July 2009 (EDT)There is a button on the software that is supposed to control the camera remotely, but while it is wired, the button is un-clickable.
23:04, 7 July 2009 (EDT)
That's good to know that it works really well wired. One really important function is the remote camera control: Taking a picture while controlling and viewing from the computer. Does this work well over the wired?
Linh N Le 01:08, 8 July 2009 (EDT)There is a button on the software that is supposed to control the camera remotely, but while it is wired, the button is un-clickable.
  • Koch wants to use the camera as a gel camera.
  • He wonders how well it works wired
    • The answer to that is very well
  • The camera can dump all the photos it has taken with the push of one button
    • The destination folder on Larry's computer is in the C: drive
    • It also sub categorizes the photos by date taken
  • As for a long usb cable, Im pretty sure they make em almost as long as you would ever want
    • The camera uses usb mini-b, so we may want to get an adapter (the thought being get a really long usb A cord and get an A to mini b adaptor, that way the long cord can be used for other things as well)
  • Amazon has usb A extension cables (15-20ft) for under $10
  • So Koch ordered two Canon Cameras and wants me to try to get the second one setup

Round 2

  • I am uninstalling the software from last time b/c I had trouble installing it the first go
    • Actually, having a hard time uninstalling the software...stupid Canon
  • Ok, all uninstalled...lets see if it will let me reinstall all the software
    • Had a problem yesterday of installing and then the program crashed. It must have left some residue behind since it says that everything is all nice and installed, yet it was not there
      • The "Photostitch" program is buggy. It tells me that it has already been installed, then crashes the installer when it fails. I can get around this by intalling manually and then telling it not to mess with photostitch
  • It seems that I have the "essential" software installed
  • Looks like it does not come with a memory card, hope thats ok
    • The new battery is uncharged, using one from old cam
  • Setting up wireless
    • SSID Kochlab
    • security WPA-PSK-TKIP
    • key Andy Maloney 00:17, 8 July 2009 (EDT): Really? You put the key on the public wiki? I know we are an open lab but don't you think that's a bit much?
      • Linh N Le 01:07, 8 July 2009 (EDT)Oh shoot! I'm sorry, i totally spaced that I did that. I just wanted to document what I wrote down so that if the camera didn't connect it could have been traced to a bad password. I gotta keep reminding myself that this is a public wiki
  • Got that setup but when trying to connect this battery also died
    • May need to wait for a charge tomorrow

Kinesin Activity Assay

  • Brigette wanted to try to do the phosphorous assay kit that we got from cytoskeleton
  • From what I know about this, we mix up tubes and ATP and let it sit at different times and measure the phosphorus levels
  • We then mix our kinesin and the tubes and ATP, let sit at diff times, and re measure the phosphorus
  • Right now, we are going to polymerize some non-labeled tubes
  • Brigette is writing up some procedures which I hope to copy from her and paste in my notebook

Procedure

  • Taking note of what we did as we do it:
  • 1 aliquot of the unlabeled tubes
    • They are in the -80c in the large falcon tubes
  • Polymerize them for 20min @ 37C
    • Add 99ul of BRB80T for a total of 100 ul of tubulin
      • note: the BRB80T is cold in the 4c fridge, so i warmed the tube by hand for ~5mins
    • Tubes + brb80t created at 12:10pm
  • Brigette is making a little bit of everything to check and see if any of it ends up killing the kinesin
    • We are doing just tubes, just kinesin, tubs + kinesin, the motillity assay with casein, etc
  • The idea here is to make different kinds of samples and let them all sit for about the same amount of time (~30mins)
    • Samples to be made:
      • 30ul MT + 30ul Kin
      • 30ul MT + 30ul Kin + .6ul ATP (1uM ATP)
      • 30ul MT + 30ul BRB80T + .6ul ATP (1uM ATP)
      • 30ul MT + 30ul Kin + 1.2ul ATP (2uM ATP)
      • 30ul MT + 30ul BRB80T + 1.2ul ATP (2uM ATP)
      • Note: When i say MT I mean MT+AF (motillity soln w/o casein)
      • Second Note: some drops got stuck to the side walls of the tubes, so Brigette washed them down with a couple ul of BRB80
  • We let the samples sit for ~10mins to allow the reactions to complete (since adding the dye and quencher stop the reactions all together)
  • Turns out it is over diluted and Brigette didn't get any useful readings from it

Trial 2

  • We are redoing it, but w/o any naked brb80
  • Samples:
      • Motility Assay alone
      • Mot +1uM ATP
      • Mot +2 ATP
      • Mot+ 3 ATP
      • Mot
      • Mot+Kin
      • Mot+kin+ATP
      • Mot+kin+2ATP
      • Mot+Kin+3Atp
  • Allowing it to sit and react for ~ 30mins
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