Team:USTC CHINA/Notebook/Protocols/Plac promoter response to IPTG or Glucose

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Revision as of 02:10, 26 September 2013

Plac promoter response to IPTG or Glucose

1. Streak a LB plate of the strain. 2. Inoculate two 3ml cultures of supplemented M9 Medium and antibiotic with single colony from the plate.
3. Cultures were grown in test tubes for 16hrs at 37℃ with shaking at 200rpm.
4. Cultures were diluted 1:1,000 into 3ml fresh medium and grown for 4.5hrs.
5. Stock concentration of the IPTG or glucose (the concentration of the both stock solution is 0.1 M) is diluted and added to different tubes to yield different final concentrations.To ensure the same response time , the IPTG or glucose should be added with a time interval of 2mins between tubes, so do the measurements procedure.
6. Measure the fluorescence (performed with SpectraMax M5 Multi-Mode Microplate Reader，add 200ul liquid in each well) and absorbance (HITACHI UV-VIS spectrophotometer U-2810 ,200ul quartz cell,path length 10mm,600nm,1.5 nm slit width) every 30 minutes in the next 4hrs.

Notebook

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