Lissa1: July17-July26: Difference between revisions
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#ImageQuant | #ImageQuant | ||
#Transform SSY403GHC into pGEV strain | #Transform SSY403GHC into pGEV strain | ||
#Continue research on Pptase optimization |
Revision as of 09:46, 24 July 2006
Monday, July 17
- Get rid of long-running SUMO gel -DONE
- Pour new SUMO gel -DONE and store -DONE
- Make new media SCD-trp+G418 -DONE
- Pour new SCD-u-t plates -DONE
- Make media for nocodazole arrests -DONE and refrigerate -DONE
- Try to figure out unpolymerized acrylamide leftovers -DONE, 200 uL TEMED added
Tuesday, July 18
- Do native extraction from blue-circle freezer yeast -DONE
- Do phosphotase protcol -DONE
- Load SUMO gel -DONE
- Set up 3 overnights: ACYL379 (in YEPD), Fus3delta (in SCD-trp), Fus3delta/Kss1delta (SCD -trp + G418). -DONe
Wednesday, July 19
- Arrive early! -DONE!
- Pour 2 new small gels -DONE and store at 4 degrees -DONE
- Grow up and flash freeze the samples for running the 2 small gels -DONE -(including pheromone arrests) -DONE
- Set up 60 mL overnight of 403 in SCR-u-h. -DONE
- Group meeting! -"DONE"
Thursday, July 20
- Cut, transfer,-DONE set up overnight of pptase gel, pour new gel -DONE
- Prep samples, -DONE load, and run -DONE the 2 small Westerns, and transfer, -DONEset up overnight
- Nocodazole arrest! -DONE
- Make a plate of the yeast ACL sent
Friday, July 21
AM:
- Wash and image all 3 Western blots
- Prep and load the nocodazole samples
PM: Lab Treat
Weekend, July 22-23
- Saturday: Lab Treat
- Sunday: Set up transfer + overnight of SUMO gel - BUFFERS HAD RUN OUT SO GEL WAS ONLY HALF-RUN. MADE NEW BUFFERS OF BOTH KINDS, REPLACED IN GEL APARATUS AND CONTINUED RUNNING.
Monday, July 24
- Call our friend Jake (ext 122) and get some Luminol.
- Learn how to use fluorescence microscope to look at induced cells
- Learn exactly what is in ACL's strains
- Set up overnight of the appropriate strain with pGEV in it
- Learn how to do phenol DNA extraction
- Cut, transfer, overnight incubation of noc. arrest gel, best done in PM
- Start to set up model of PPL system on paper
- Research ways to debug Pptase gel
- Add buffer to CIP reaction?
- Increase amount of PPTase?
- Try another PPtase?
- Look at NEB catalog
- Look at other papers that have done Pptase
Tuesday, July 25
- Phenol DNA extraction
- Set up PCR of extracted DNA with the forward and reverse primers I ordered
- ImageQuant
- Transform SSY403GHC into pGEV strain
- Continue research on Pptase optimization