Imperial College London/Notebook/16 September 2009

Testing

Lac characterisation 16th Sep

Protocol considerations

• Overnight cells were diluted in fresh media 1:12
• The cells were allowed to grow until their OD reached around 0.6 (around 4 hours)
• IPTG was added from a stock solution of 1M IPTG and 0.1M IPTG. Note that IPTG takes some time to defrost
• 180ul of Media with relevant IPTG conc was added to each 96 well plate. 20ul of cells were added.
• Put in the plate reader overnight using protocol IGEM Fluor Abs (do not use)

Results

Download raw data

Conclusions

• There was no change in the cell growth rate with IPTG
  • Since a Lac-RFP construct was used, this shows that IPTG, for both its toxicity and protein production stress, does not have much effect on cell growth at the concentrations tested

Suggestions/Improvements

• The script merged the fluorescence readings into the absorbance readings.
  

After overnight readings, the regular absorbance readings were obtained, but no fluorescence readings were obtained. Therefore, the results had to be converted to a normal cell growth analysis.

• • Try to give the script test runs before leaving them overnight, and check on results regularly if possibly