Lissa1: July27-August2

Wednesday, July 27

1. Run a gel of the PCR reaction and cut out the pGEV band. - DONE, but pGEV was not visible
2. Pour little gels and debug gel-casting system, store -DONE
3. Run a new pCR of ACLY700 and YAS with more controls overnight -DONE
4. Put in new overnights (again) of parent, Fus3delta, Fus3Kss1delta strains. -DONE
5. Put in LARGE overnight for nocodazole arrests (403 in SCR-u-h), and another one to make more blue-circle samples. -DONE
6. Make media for nocodazole arrests - DONE

 TY'S plate!!!!!!!!!!!!!!! -DONE

Thursday, July 28

1. Run a new gel of the PCR from yesterday -DONE
   1. Run yet another gel of yesterday's PCR -DONE
2. Beta-estradiol stocks. - DON'T KNOW HOW!!!
3. Plan induction experiment -DONE
4. Prep proteins (AM) -DONE, but it took them *forever* to grow.., load and run gels, set up transfer - NOT FEASABLE
5. Nocodazole arrest, freeze samples. - DONE
6. Gel extraction - DONE
7. Make buffers -DONE
8. Alpha-arrests -DONE

Friday, July 29

1. Induction experiment using pGEV system - SADLY, DOES NOT EXIST YET
2. Pour new SUMO gel -DONE
3. Do another phosphatase/native extraction prep (with more CIP, buffer, etc), and load on a new SUMO gel.
4. IMAGEQUANT

Weekend, July 30-31

1. Saturday - will be in Western Massachusetts
2. Sunday - will be sailing - evening: set up transfer of phosphatase gel.

Monday, August 1

1. Pour new SUMO gel, prep nocodazole samples and load.
2. Long incubation, washing and imaging of Western blot.
3. Prep samples, load and run 2 small gels, transfer, incubate.