Lissa1: August6-August14

August 7

 * 1) Pour new SUMO gel -DONE
 * 2) Pour new small gels -DONE
 * 3) Set up overnights for the following experiments:
 * 4) Nocodazole arrest -DONE
 * 5) Induction experiment -DONE
 * 6) Make more unarrested 403 for phosphatase Western -DONE
 * 7) Make media for the nocodazole arrest. -DONE
 * 8) Plan EVERYTHING -CHECK
 * 9) Cloning
 * 10) Finalized experiments
 * 11) What to do about luminol/ECF deal? - OOPS, stil don't know
 * 12) Integrate new-found numbers/data in to model -DONE
 * 13) Maybe update model to include Michaelis-Menton kinetics -NOT YET, talk to Ty

August 8

 * 1) Do nocodazole arrest! -CELLS WON'T GROW:(
 * 2) Do induction experiment! -DONE
 * 3) Set up overnights for the following experiments: -MOVED UNTIL TOMORROW
 * 4) Fus3/Active Fus3 gels
 * 5) Phosphatase control
 * 6) PCR up pGEV out of ACLY700 -DONE
 * 7) Run a gel of the PCR -DONE
 * 8) If there's time, run a second gel and extract and purify the pGEV (which might not be pGEV, based on the sequencing data) -DONE

August 9

 * 1) PCR amplify the pGEV band I purified yesterday -DONE
 * 2) PCR out GEV insert from the original pGEV DNA -DONE
 * 3) Run gels of these PCRs and do gel extraction -DONE
 * 4) Digest pGEV - no time
 * 5) Setup overnight of pRS-405 in DH5alpha, in LB + Amp -DONE
 * 6) Streak new plate -DONE
 * 7) Setup overnight of samples for Fus3, etc -DONE


 * Unfortunately, I've got a cold and need to rest:(

August 10

 * 1) Prep Fus3 samples (all the way to sample buffer) and store -DOne
 * 2) Get concentrations on all DNA - DONE
 * 3) Miniprep 405 out of E. coli -DONE
 * 4) Streak out new 403 plate from freezer stock -DONE
 * 5) Digest pGEV, run on gel to check -DONE
 * 6) Design diagnostics for prs405 DNA -DONE
 * 7) Digest 405 -DONE
 * 8) Do PCR cleanup - DONE

August 11

 * 1) Make new SCR-u-h media!
 * 2) Do native extraction and phosphatase experiment on Noc. arrested yeast
 * 3) Load SUMO gel and start run
 * 4) PCR up GEV insert out of miniprepped pGEV DNA

August 12

 * 1) Gel-extract and purify all digests/PCRs. -DONE
 * 2) Check on SUMO gel l- SOMETHING IS VERY WRONG. CHANGED ALL BUFFERS, CHECKED ALL CONNECTIONS, SET UP AGAIN.  DID NOT SEE BUBBLES AT ALL.  1 HR LATER IT HAD MOVED A LITTLE BIT....

August 13

 * 1) The plan was to set up the transfer of my gel today, but whatever problem was present yesterday is also present today. I think a wire must have snapped inside the gel box?  It's giving 400 V but only 2 mA of current...

August 13, 2006

 * 1) Digest my new PCR-ed fragments that hopefully are pGEV with the double digest -DONE
 * 2) At the end of the day, do the ligation
 * 3) Think about this week's very complicated schedule -DONE
 * 4) Set up overnight of my newly-streaked-out 403 cells to do noc. arrest and also to set up new samples for phosphatase control -DONE
 * 5) Get opinions on induction experiment -DONE
 * 6) Set up new overnight for another induction
 * 7) Scrap old SUMO gel and pour a new one
 * 8) Prep and run 2 new small Fus3/etc gels -DONE