IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/24

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QS Track

  • Like the dspB track, a bacterial mat grew when transformants were spread on chloramphenicol plates.
  • A bacterial mat also grew when transformants were spread on LB + 20 μL chloramphenicol (1000X).
  • This suggests a possible problem with the chloramphenicol aliquots (1000X) used.
  • Marianne remade the chloramphenicol aliquots using 3 different sources of chloramphenicol.
  • Phillip made chloramphenicol plates (20) using Marianne's C3 aliquots (chlor source: last year's parafilm sealed bottle)
  • From yesterday's leftover ligation mixtures, 5 μL was transformed again by Vicki (following UBC iGEM procedures).
  • Also started an overnight culture of pcN33. Host is Staph aureus RN4220. Used LB broth (no erm in lab).
  • Made an antibiotic test of possible problematic chloramphenicol aliquots mentioned earlier. 10 μL of chloramphenicol aliquot was added to 10 mL LB. A stab of stock DH5(alpha)was inoculated in this medium. Expect no growth.

dspB Track

Gel verification on RD of chlor & amp backbones

  • Protocol: SOP
    • Changes: 0.08% agar
  • Machine conditions: 0.5x TBE buffer, 100V, 60 min

Gel orientation:

Gel orientation
1 kb ladderChlor EChlor XChlor P100 bp ladderAmp EAmp XAmp P1 kb ladder

Results:

Transformation on ligation mixes from Aug 13 2010

  • Ligation mixes: 1,4,5,8,11,14,15,AA,TB,his, no-his
  • Protocol: Common protocol - SOP
    • Changes: add 2uL of ligation mixes instead of 1uL
  • Transformants put in 37C incubator @ 1630

O/N Culture for making competent cells

  • Put in at 1630


Biofilm Track

Eric F. and Melody

Biofilm Growth Protocol Day 2 (100823E + M)

  • O/N cultures removed from incubator @ 1030 - 20 hours in incubator
  • Control showed no growth
  • Plate Layout
=100823E + M
Row 3 4 5 6 7 8 9 10
C 8 C R C 8 C R C
D 8 C R C 8 C R C
R 8 C R C 8 C R C
  • Note:
    • R = RN4220 + 1% Glucose TSB
    • 8 = 8325 - 4 + 1% Glucose TSB
    • C = control + 1% Glucose TSB



  • Any contamination that occurs in C8 (100823E) is isolated
  • 100823E into incubator @ 1056
  • 100823M into incubator @ 1305

Biofilm Growth Protocol Day 1 (100824E + M)

  • Followed protocol
  • RN4220 + 8325-4 + Control
    • 5mL TSB each
  • Into incubator @ 1500



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