Lissa1: July17-July26

Monday, July 17

 * 1) Get rid of long-running SUMO gel -DONE
 * 2) Pour new SUMO gel -DONE and store -DONE
 * 3) Make new media SCD-trp+G418 -DONE
 * 4) Pour new SCD-u-t plates -DONE
 * 5) Make media for nocodazole arrests -DONE and refrigerate -DONE
 * 6) Try to figure out unpolymerized acrylamide leftovers -DONE, 200 uL TEMED added

Tuesday, July 18

 * 1) Do native extraction from blue-circle freezer yeast -DONE
 * 2) Do phosphotase protcol -DONE
 * 3) Load SUMO gel -DONE
 * 4) Set up 3 overnights: ACYL379 (in YEPD), Fus3delta (in SCD-trp), Fus3delta/Kss1delta (SCD -trp + G418). -DONe

Wednesday, July 19

 * 1) Arrive early! -DONE!
 * 2) Pour 2 new small gels -DONE and store at 4 degrees -DONE
 * 3) Grow up and flash freeze the samples for running the 2 small gels -DONE -(including pheromone arrests) -DONE
 * 4) Set up 60 mL overnight of 403 in SCR-u-h. -DONE
 * 5) Group meeting! -"DONE"

Thursday, July 20

 * Cut, transfer,-DONE set up overnight of pptase gel, pour new gel -DONE
 * 1) Prep samples, -DONE load, and run -DONE the 2 small Westerns, and transfer, -DONEset up overnight
 * 2) Nocodazole arrest! -DONE
 * 3) Make a plate of the yeast ACL sent

Friday, July 21
AM: PM: Lab Treat
 * 1) Wash and image all 3 Western blots
 * 2) Prep and load the nocodazole samples

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

 * 1) Call our friend Jake (ext 122) and get some Luminol. -DONE, but no luminol
 * 2) Learn how to use fluorescence microscope to look at induced cells
 * 3) Learn exactly what is in ACL's strains
 * 4) Set up overnight of the appropriate strain with pGEV in it, and the parent strain
 * 5) Learn how to do phenol DNA extraction
 * Cut, transfer, overnight incubation of noc. arrest gel, best done in PM -DONE
 * 1) Start to set up model of PPL system on paper -DONE
 * 2) Research ways to debug Pptase gel -DONE
 * 3) Add buffer to CIP reaction? -GOOD IDEA
 * 4) Increase amount of PPTase? -?
 * 5) Try another PPtase? -THIS SHOULD WORK
 * 6) Look at NEB catalog - Calbio has really nice phosphatases
 * 7) Look at other papers that have done Pptase -DONE

Tuesday, July 25

 * 1) Wash and image Western blot -DONE
 * 2) Phenol DNA extraction -DONE
 * 3) Set up PCR of extracted DNA with the forward and reverse primers I ordered-DONE
 * 4) Also try integration verification primers from previous pGEV integration attempts. -DONE
 * 5) ImageQuant -MOVED because people were using the computer by the typhoon
 * 6) Transform SSY403GHC into pGEV strain and parent strain (starting from beginning of EZ trans)- by the time I got around to this part, my cultures had overgrown for a second time (they had overgrown while I was doing the Phenol DNA extraction) and I couldn't prepare the competent cells. I'll have to do this some other day.
 * 7) Make freezer stock of Alejandro's strains. - equal parts yeast and glycerol, slow freeze -DONE
 * 8) Size comparison of Kss1 and Fus3. Does it make sense to run them on a SUMO gel? -DONE, no