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.
4. Put in overnight of ACLY700.

Tuesday, August 2

1. Long incubation and imaging of Fus3 Western -DONE
2. Transform 403 in to ACLY700 and plate. -DONE
3. PCR ACLY700 DNA, run a gel, gel extract, send for sequencing. -DONE
4. SCBT (probably tomorrow)
5. Order spacer plates. -DONE
6. Math model. -THINKIN'

Wednesday, August 2

1. Cut, transfer of nocodazole gel.
2. Call SCBT? - As of yesterday no longer seems like a good idea
3. Pour 2 small gels. -PENDING...
4. Run a new set of Fus3/activeFus3 gels, set up transfer.
5. Model -NEED PARAMS

Thursday, August 4

1. Wash and image Nocodazole gel
2. Long incubation, wash, image gel from Weds.
3. Get cells of ACLY700-403 plates, grow up overnight.
4. Figure out beta-estradiol stocks.

Friday, August 5

1. Beta-estradiol stocks.
2. Induction experiment.