Wednesday, July 27
- Run a gel of the PCR reaction and cut out the pGEV band. - DONE, but pGEV was not visible
- Pour little gels and debug gel-casting system, store -DONE
- Run a new pCR of ACLY700 and YAS with more controls overnight -DONE
- Put in new overnights (again) of parent, Fus3delta, Fus3Kss1delta strains. -DONE
- Put in LARGE overnight for nocodazole arrests (403 in SCR-u-h), and another one to make more blue-circle samples. -DONE
- Make media for nocodazole arrests - DONE
TY'S plate!!!!!!!!!!!!!!! -DONE
Thursday, July 28
- Run a new gel of the PCR from yesterday -DONE
- Run yet another gel of yesterday's PCR -DONE
- Beta-estradiol stocks. - DON'T KNOW HOW!!!
- Plan induction experiment -DONE
- Prep proteins (AM) -DONE, but it took them *forever* to grow.., load and run gels, set up transfer - NOT FEASABLE
- Nocodazole arrest, freeze samples. - DONE
- Gel extraction - DONE
- Make buffers -DONE
- Alpha-arrests -DONE
Friday, July 29
- Induction experiment using pGEV system - SADLY, DOES NOT EXIST YET
- Pour new SUMO gel -DONE
- Do another phosphatase/native extraction prep (with more CIP, buffer, etc), and load on a new SUMO gel.
Weekend, July 30-31
- Saturday - will be in Western Massachusetts
- Sunday - will be sailing - evening: set up transfer of phosphatase gel.
Monday, August 1
- Pour new SUMO gel, prep nocodazole samples and load.
- Long incubation, washing and imaging of Western blot.
- Prep samples, load and run 2 small gels, transfer, incubate.
- Put in overnight of ACLY700.
Tuesday, August 2
- Long incubation and imaging of Fus3 Western -DONE
- Transform 403 in to ACLY700 and plate. -DONE
- PCR ACLY700 DNA, run a gel, gel extract, send for sequencing. -DONE
- SCBT (probably tomorrow)
- Order spacer plates. -DONE
- Math model. -THINKIN'
Wednesday, August 2
- Cut, transfer of nocodazole gel.
- Call SCBT? - As of yesterday no longer seems like a good idea
- Pour 2 small gels. -PENDING...
- Run a new set of Fus3/activeFus3 gels, set up transfer.
- Model -NEED PARAMS
Thursday, August 4
- Wash and image Nocodazole gel
- Long incubation, wash, image gel from Weds.
- Get cells of ACLY700-403 plates, grow up overnight.
- Figure out beta-estradiol stocks.
Friday, August 5
- Beta-estradiol stocks.
- Induction experiment.