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.

Gel verification on RD of chlor & amp backbones
Gel orientation: Results:
 * Protocol: SOP
 * Changes: 0.08% agar
 * Machine conditions: 0.5x TBE buffer, 100V, 60 min

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 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

 * 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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