IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 1.1

Results summary

Experiment 1

• Concentration of pTet-GFP = 110ng/ul
• Concentration of pT7-GFP = 100ng/ul
• Concentration of pCI-GFP =

Experiment 2

• pTet-GFP has been successfully tested in vivo. The levels of fluorescence of the construct where comparable to the levels of our positive control (200x dilution of GFP) meaning that GFP was actually produced. Therefore pTet-GFP works in E.Coli.

• pT7-GFP testing failed because it was not properly induced. It will be re-tested on the 17/08/2007 after it has been induced with IPTG and left overnight to allow for a measurable GFP amount to be produced. IPTG is a T7 inducer for the E.Coli strain used (BL21).

pT7 was tested the next day and no fluorescence was observed. Two reasons were hypothesized: The bacteria cells were in stationary phase; or that the pT7 construct is not working.

• pCI-GFP testing has been postponed due to delays in the making of the construct. Testing will be carried out as soon as the construct is ready.

• A 1 hour run (taking measurements every 2.5 min) was also conducted to test the variation on the expression levels of pTet-GFP over time.

Results of testing pTet and pT7 at 0.15 sec counting time for fluorometer detector

Results of testing pTet and pT7 at 0.15 sec counting time for fluorometer detector

Complete set of results and raw data .