From 2007.igem.org

Objective:

check if our lux-lac hybrid promoter is activated by AHL

Samples:

Lux-lac hybrid promoter (before blue light check*) x3 samples
Lux-lac hybrid promoter (after blue light check) x3 samples
placQI (pos. con.)
modified A4 (promoter deleted) (neg. con.)

(*Blue light check: Transformed with the hybrid promoter + GFP, the colonies should look green irradiated by blue light. Greenish colonies were picked up for the overnight culture. In addition, since this irradiation could affect the cells and their genes, colonies before the irradiation were picked up without confirming if they looked green. )

Procedure:

AHL assay Standard protocol

Result & Conclusion:

2 out of the 3 samples of Lux-lac hybrid promoter (before blue light check*) and
1 out of the 3 samples of Lux-lac hybrid promoter (after blue light check*)
showed significant fluorescence; whereas,
1 out of the 3 samples of Lux-lac hybrid promoter (before blue light check*) and
2 out of the 3 samples of Lux-lac hybrid promoter (after blue light check*) did not.
1 out of the 3 samples of Lux-lac hybrid promoter (before blue light check*) showed weak fluorescence.
(For the last sample, we detected that this sample emerged from cells from two colonies, one containing the hybrid promoter + GFP and the other not by mini-prep and electrophoresis.)

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