IGEM:Imperial/Results/S01656

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S01656 Protocol Page

Sunday, 20 August 2006

  • Normalised Fluorescence of +IPTG +AHL
Tube 1 - 233
Tube 2 - 237
Tube 3 - 239
  • Normalised Fluorescence of +IPTG -AHL
Tube 1 - 219
  • Normalised Fluorescence of -IPTG +AHL
Tube 1 - 244
  • Normalised Fluroescence of -IPTG -AHL
Tube 1 - 218
  • Comparison between +IPTG +AHL and -IPTG +AHL:
% Difference = 2.94%
Conclusion: Not statistically significant
  • Resolution
Test expression of AiiA protein by SDS electrophoresis
Re-test the protocol on Tuesday 22 August 2006

Tuesday, 22 August 2006

  • Different formulation used to calculate normalised fluorescence. Used the same calculations as the T9002. See updated spreadsheet.
  • Normalised Fluorescence of +IPTG +AHL
Tube 1 - 4409
Tube 2 - 4659
Tube 3 - 4574
  • Normalised Fluorescence of +IPTG -AHL
Tube 1 - 4574
  • Normalised Fluorescence of -IPTG +AHL
Tube 1 - 4769
Tube 2 - 4762
Tube 3 - 4834
  • Normalised Fluroescence of -IPTG -AHL
Tube 1 - 4445
  • Comparison between +IPTG +AHL and -IPTG +AHL:
% Difference = 4.25%
Conclusion: Not statistically significant
  • Resolution
Test expression of AiiA protein by SDS electrophoresis
Re-test the protocol on Wednesday 23 August 2006 with higher concentration of AHL

Wednesday, 23 August 2006

Results of western Blot

Image:Non-working Gel.JPG

The IPTG has not induced the production of a protein which weighs 28736 kd (aiiA) this should be loacted inside the oval. We are repeating the expperiment to confirm the results. We are sequencing the part to confirm the results. The ladder is invitrogen marker 12.

JohnChattaway 10:13, 23 August 2006 (EDT)

Results of AHL Testing

  • Used 100 μM stock concentration instead of 10 μM. Used 800 μL of supernatant and 1200 μL of T9002 cells.
  • Normalised Fluorescence of +IPTG +AHL
Tube 1 - 4353
Tube 2 - 3552
Tube 3 - 4476
  • Normalised Fluorescence of +IPTG -AHL
Tube 1 - 4157
  • Normalised Fluorescence of -IPTG +AHL
Tube 1 - 3688
Tube 2 - 4609
Tube 3 - 4197
  • Normalised Fluroescence of -IPTG -AHL
Tube 1 - 3608
  • Comparison between +IPTG +AHL and -IPTG +AHL:
% Difference = 0.91%
Conclusion: Not statistically significant
  • Resolution
Re-test the protocol on Thursday 24 August 2006 with same concentration of AHL
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