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:
   1. Nocodazole arrest -DONE
   2. Induction experiment -DONE
   3. Make more unarrested 403 for phosphatase Western -DONE
4. Make media for the nocodazole arrest. -DONE
5. Plan EVERYTHING -CHECK
   1. Cloning
   2. Finalized experiments
   3. What to do about luminol/ECF deal? - OOPS, stil don't know
6. Integrate new-found numbers/data in to model -DONE
   1. 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
   1. Fus3/Active Fus3 gels
   2. Phosphatase control
4. PCR up pGEV out of ACLY700 -DONE
5. Run a gel of the PCR -DONE
6. 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