Lissa1: July1-July8

Weekend

 * 1) Wash and image Western on Saturday morning

July 3, 2006

 * 1) Start SUMO gel with ATP analog/arrested yeast from Friday. - DONE
 * 2) SO FEW YEAST - is something wrong? What protocol to use?  Same one?
 * 3) Who is "the boy who lived" on my pGEV transformation?
 * 4) Work on biobricking PPL parts (now that I have Endy Lab membership)
 * 5) What if the pGEV we have is on a CEN/ARS plasmid with a Trp marker, like the paper says? Is this even a remote possibility?
 * 6) Come up with schedule for rest of week... July 4 vacation day? - DONE
 * 7) Colony PCR of "the boy who lived" - DONE
 * 8) Transform new pGEV in to E. coli - DONE

July 4, 2006

 * 1) Day off? Institute holiday...

July 5, 2006

 * 1) Native extraction! using unarrested 403 - DONE
 * 2) Pour new sumo gel for native extraction samples - DONE
 * 3) Transfer and overnight incubation of Western from July 3 - DONE
 * 4) Run a gel of the colony PCR from Monday - DONE
 * 5) If it works, and pGEV is there, grow up overnight culture from new plate
 * 6) If it works, and pGEV isn't there...
 * 7) If it doesn't work, try again (see July 6).
 * 8) Conclusion: something is happening, but it isn't either bad nor good.  Mixed bag.  Try again.
 * 9) Order anti-Fus3 antibody from Santa Cruz, order Rainbow Marker from GE Healthcare. - DONE
 * 10) Set up overnight culture of pGEV E. coli in LB+Amp. - Had to plate anew (1:200) instead, but that is DONE
 * 11) Set up overnight of 403-1030-1018 in SCD -u -h. -DONE

July 6, 2006

 * 1) Wash and image Western - DONE
 * 2) Try colony PCR again (AM) and run gel - DONE
 * 3) Grow up overnight culture of pGEV404 in DH5alpha in LB+Amp. -DONE
 * 4) Grow up overnight culture of 403-1030-1018 in SCR -u -h. - DONE
 * 5) Set up freezer stock (the kind in glycerol) of 403-1030-1018 from yesterday's overnight. -DONE
 * 6) Debug peristaltics - DONE

July 7, 2006

 * 1) Miniprep pGEV out of E. coli, do characteristic digest (run gel), linearize pGEV with SnaBI (run gel). -DONE
 * 2) Transfer and overnight incubation of Western - set up transfer
 * 3) If sumo gel with noc-arrested samples didn't work, re-arreast cells. - DONE, even though it worked
 * 4) If there's time, do a PCR of the yeast genome with Trp1_A and Trp1_D.- MOVED TO MONDAY
 * 5) Transform pGEV in to 1030-1018 using green star EZ method - DONE

July 8, 2006

 * 1) Set up overnight incubation of Western - DONE

July 9, 2006

 * 1) Imaging of Western -DONE
 * 2) Pour new 12% SUMO gel - DONE
 * 3) Prep arrested cells from Friday and load on gel - DONE