IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/07/27

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Quorum Sensing Track

Miniprepped DH5(alpha) cultures containing 34 (E0840), 35 (I13504), and 36 (J74107) following Raf's Alkaline Lysis protocol (link:).

Digested 34 and 35 with XP following UBC iGEM digestion protocol (link:).

Digested P2 promoter (I746104), J23100, and Pbad promoter (wildtype: I13453) using ES following UBC iGEM digestion protocol.

Digested PSB1C3 plasmid with EP following UBC iGEM digestion protocol.

  • Ligate the above 3 kinds of promoter (and another medium transcription promoter: I14033) with 34 and 35. This results in a total of 8 different combinations. Follow UBC iGEM ligation (6:1 insert to vector ratio) protocols and leave overnight*

QS Track

Jason and Eric F.

SH1001 Growth Curve

Used growth curve protocol found in the wave titled "BioFilm Track Pt2". Procedure and SOP will be uploaded to OWW in the future.

These notes begin on Day 2 of the growth curve experiment

  • Have SH1001 overnight culture w/ no growth in the control
  • Will do 3 experiments today with different initial dilution factors
    • 1/200
    • 1/300
    • 1/400
  • Previous experiments with this strain using the procedure's dilution factor (1/25) gave OD600 readings that were too high (greater than 0.4)
  • Total volume for these experiments: 15mL
Control 1/200 1/300 1/400
B2 Broth (ml) 15 15 15 15
Overnight Culture (μL) 0 75 50 37.5



  • After the growth curves are successfully done, the cells will be diluted and grown for use as competent cells
  • Took time point 0 readings right away after gently mixing all 4 flasks


  • Into incubator for first time at 1331
    • AMBL lab incubator: set to 200 RPM - 37°C
  • Out at 1404
  • At 1404 all flasks were very bubbly


  • Back into AMBL lab incubator at 1421
  • Out at 1455
  • Bubbles still present


  • Back into AMBL lab incubator at 1521
  • Out at 1541
  • Bubbles still present


  • Into Lagally lab incubator at 1612, same settings
  • Out at 1642
  • Less bubbles, but still present


  • Experiment discontinued due to contamination of the control, see data below


Time in Incubator OD600 of 1/200 OD600 of 1/300 OD600 of 1/400 OD600 of Control OD600 of Blank
0 (a) 0.000 0.063 0.116 0.000 0.000
0 (b) 0.000 0.075 0.072 0.000 0.000
30 (a) 0.036 0.025 0.077 0.002 0.000
30 (b) 0.044 0.031 0.081 0.006 0.000
60 (a) 0.065 0.053 0.105 0.003 0.000
60 (b) 0.070 0.058 0.101 0.003 0.000
90 (a) 0.075 0.054 0.162 0.023 0.000
90 (b) 0.071 0.050 0.163 0.012 0.000
120 (a) 0.165 0.077 0.135 0.027 0.000
120 (b) 0.167 0.074 0.153 0.027 0.000




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