This is a kinetic assay for induction by HSL over a 107-magnitude concentration. Each construct is measured in triplicate.
Plates that are needed: 3+X shaker/growing, X fluorescence, where 2X is number of time points desired.
- M9+0.1%YE+antibiotic plates
- 210 mL M9+0.1%YE+antibiotic
- 96 well growing plates
- 96 well fluorescence plates
- 96 well clear bottom absorbance plates
- Condition parts on M9+0.1%YE+antibiotic minimal media plates
- Inoculate 3 single colonies from these plates into M9+0.1%YE+antibiotic media overnight
- Autoclave 96 well shaker/growing plates
- Dilute cells down to OD600=0.1
- Do not put cells on ice!
For GFP, assay at 30ºC