Arthur Yu Notebook


Revision as of 23:53, 11 June 2007 by Ayu (Talk | contribs)

My Construction Files
My Sequencing Files

6/11 austin's birthday

  • CAKE PARTY - great custard cake
  • I put the wbbL (1) and HPI/katG (4) colonies to incubate in LB broth.
  • neuS failed; no colonies :(((((((
    • redid ligation and xformation. hopefully there will be good results tmrw!
  • made like 20 LB-Agar/Amp plates - looks like our stock will last at least this week
  • researched nitric oxide (NO) and E. Coli - looks like soxRS is promising
  • also researched RBCs and how they deal with NO
  • plopped yfbE into PCR will do stuff with it tmrw

TO DO: enter yfbE into the registry
Ayu 18:24, 11 June 2007 (EDT)

6/8 long day?

  • My PCR from last night (HPI/katG) was ROXOR! (left)
    • xformed some DH10B's. w00t w00t
  • Today's PCR was wbbL and neuS. ALSO ROXOR LOL (right)
    • xformed DH10B's.
  • made oligos for yfbE promoter thingy - will test with GFP and yeah! next week!
  • poured lotsa LB/agar+amp plates

060807 ayu gel1.jpg
060807 ayu gel2.jpg

6/7 we got benches

  1. well... getting the mutated PCR prod overnight. going to xform tmrw, hope it works!
  • programmed pcr on machine upstairs (#6)
  • we got computers
  • AGAR SUX, for future reference:
  1. nuke @ 20:00 min, 50% power.
  2. water bath in tap water for 5-10 min
  3. thaw the antibiotic right now!!
  4. FIRE for disinfecting
  5. pour that stuff. set 15 min, then marker it then bag

6/6 waiting for oligos

  • Made oligos and constructs with Vai, for getting wbbL and neuS from pJ23006-Bca9106
  • We tried the P_tet/RFP triple/double digest to make a composite part.
  1. FAIL
  2. probably source DNA is bad
  3. so much for that activity...

Other stuff: I won speed scrabble. even though I kind of cheated ish (didn't stop when Sam said stop"

6/5 coolbeans

  • Finalized oligos to order with Vai
  • Learned about LB broth-ing and LB/Agar plating. Thanks, Austin and Sam :)
  • Learned about the many composite part-making methods. Props 2 Chris
  1. prefix/suffix is weaksauce
  2. Use the AlwnI or BsaI or BglI, in conjuction with BglII or BamHI << (Did this today)
  3. DBBS
  4. 3 antibiotic; MIT endorses, used for BioBrick 1.0. Triple digest = bad
  5. 1-2-3 method << 'Our Goal' in a few weeks. should be leet.
  • Planned and vicariously did the making of P_tet+RFP brick (see Vai Notebook)

Other Notes: All oligos are being ordered, w00t w00t. Ayu 18:36, 5 June 2007 (EDT)

6/4 Training Finishes, Real Stuff Starts

  • Incubated some colonies
  • Miniprep'd already-been-incubated colonies (2)
  • Double digest of the 2 minipreps + parent plasmid
  • Colony PCR'd the incubated E.coli
  • Ran gel of the digest + PCR
  • >>> PCR product / Miniprep 1 / Parent Plasmid / Miniprep 2 / Ladder >>>
  • No bands for PCR or parent. Confused? Other ones look great.

As for me: Wiki acc works now.
Designing oligos and will compare with Vai. Ayu 18:19, 4 June 2007 (EDT)

to do