IGEM:IMPERIAL/2007/Notebook/2007-10-5

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pTet-LuxR-pLux-GFP construct in-vivo

  • Aim: The aim of the test was to check if construct 1 of infector detector works in-vivo
  • The protocols for the experiment can be found here
  • The results of the test showed that pTet-LuxR-pLux-GFPmut3b is working in vivo and GFPmut3b produced increases with increasing [AHL]

pTet-LuxR-pLux-GFP construct in-vitro

  • Aim: The aim of the test was to check if construct 1 of infector detector works in-vitro, for a number of concentrations of AHL
  • The protocols for the experiment can be found here
  • The results of the test seemed very variable and didn't seemed to comply with the tests that were carried out before. There was a lot of variability between the samples and 4nM of AHL seemed to have fluorescence output greater than 50nM.
  • It was decided to carry out the experiment again. The reasons for the inaccuracies in the data could have been:
    • The reaction was not mixed properly or the right amounts were not pipetted
    • The plates had already been used and thus could have affected the reaction
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