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

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