IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/09/03

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Biofilm Track

Melody, Eric

  • Autoclaved 30ul pipette tips

Biofilm Protocol Day 4

  • 0.1% Crystal Violet stain on plates 100831M24 + 100831M26

Biofilm Protocol Day 3

  • Took plate 100901M28 @ 8:06 p.m.
  • Took plate 100901M30 @ 10:15 p.m.
    • No contamination
  • 3 x 300ul PBS wash of plate 100901M28
    • heat fixed @ 8:34 p.m.
  • 3 x 300 ul PBS wash of plate 100901M30
    • heat fixed @ 10.46 p.m.

dspB Track

  • Plates taken out at 11am
    • Lawn of colonies on all 6 plates including transformants with ccdB chlor vector. Is ccdB resistant to chloraphenicol now???

Gel verification on RD

  • Protocol: biobrick "digest" protocol
  • Changes: load ~17uL into each well; 10uL for ladder

Gel orientation:

Gel orientation
ccdb chlorbb100bp ladderRFP chlor

Machine conditions: 0.5x TBE buffer, 100V, 60min Results:

Gel extraction

  • Using Invitrogen PureLink Quick Gel Extraction Kit (protocol inside kit)

ccdB chlor gel weight: 0.0790g
RFP chlor gel weight: 0.0878g

Nanodrop:

  • [ccdB chlor] = 5.5ng/uL
  • [RFP chlor] = 5.2ng/uL

Transformation

  • Protocol: SOP
  • Put in 37C incubator @ 6:47pm
  • Plates:
      1. 1: ccdB no his
      2. 2: ccdB his
      3. 3: ccdB no his Alina
      4. 4: ccdB his Alina


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