5 September 2007

Construction of pT7-GFP

1. Innoculated 5ml of LB + Kan with 550μl of fresh overnight culture
2. Grew culture in 37°C until O.D.600 = 0.6
3. Induced expression of RNA polymerase with 1mM IPTG
4. Checked for fluorescence after 4 hours of induction
5. Colony 1 showed a fluorescence reading of ~200 000
   • Negative control of non-induced Colony 1 had a reading of ~3 000
6. Colony 4 showed a fluorescence reading of ~3 000
   • Negative control of non-induced Colony 4 had a reading of ~3 000
7. Colony 1 seems to contain the correct pT7-GFP DNA construct

Testing of different [AHL] on pTet-LuxR-pLux-GFP DNA construct

[AHL] of 3nM, 5nM and 7nM was tested. Results can be found here.

Protocol can be found here under Prelimineary AHL testing on the experimental design page.

Results can here under Results on the experimental design page.