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
2. At the end of the day, do the ligation
3. Think about this week's very complicated schedule
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
5. Get opinions on induction experiment
6. Set up new overnight for another induction