IGEM:Imperial/2007/Wet Lab/results/CBD4

= Testing DNA concentration of pTet-GFPmut3b In vitro=

Aims
To determine if the optimum concentration of pTet-GFPmut3b in vitro''.

Materials and Methods
Link to Protocols page

Results
Controls:
 * Negative Control- Nuclease Free Water was added instead of DNA

Constants:
 * Temperature - 37&deg;C
 * Total Volume - 60&micro;l

Raw Data
 * [[Media:IC 2007 CBD DNA Concentration.xls|Raw Data excel file]]

Discussion
Figure 1.1 and Figure 1.2. show us that the synthesis of GFPmut3b increases with DNA added. Though the molecules of GFPmut3b synthesised increases with increasing DNA concentration, even after a concentration of 4&micro;g. This is not what was expected because of the nature of the cell free chassis which is optimum DNA concentration = 4&micro;g. But the increase in fluorescent moecules synthesised by increasing DNA concentration is not very big. If DNA concentration is increased from 2&micro;g to 6&micro;g (a percentage increase of 200%), it reuslts in only about 15% increase (at 360min) in GFPmut3b molecules synthesised. Thus it was decided that a DNA concentration of 2 or 4&micor;g will be used for testing pTet-GFPmut3b construct.

Conclusion
To conclude the following approximations can be made:

Optimum DNA concentrations - 2 or 4µg of DNA