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

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Media test

  • The aim of this test was to see if the fluorescence of GFP mut3b gets affected by the media it is in.
  • We tested the reporter's expression in nuclease free water and LB media and then compared it with the reults got from GFP mut3b calibration curve
  • The results can be found here
  • It was found a linear relationship between the fluorescence of GFP mut3b in the three media.
  • This meant that LB media could be used to get calibration curve for dsRED express instead of cell extract. But as dsRED express couldn't be cloned, we didn't make a calibration curve for dsRED express

pT7 test in-vitro

  • The aim of the test was to see if pT7-GFP mut3b and pT7-dsRED express work in-vitro or not
  • The protocols can be found here
  • The experiment didn't give good results as:
    • The incubator had been turned off, thus the samples were not at the right temperature.
    • The right filter for dsRED express was not used to begin with
  • After using the right filter, it was realised that pT7-dsRED express construct doesn't express
  • The results can be found here
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