Team:Aachen/Project/2D Biosensor
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=== Detecting 3-oxo-C{{sub|12}} HSL with sensor chips === | === Detecting 3-oxo-C{{sub|12}} HSL with sensor chips === | ||
- | {{Team:Aachen/FigureFloatRight|Aachen_K131026_Platereader.gif|title=Testing K131026 in our sensor chips|subtitle=K131026 in our sensorchip induced with 0.2 µl 3-oxo-C{{sub|12}} HSL and measured with a Platereader. Blue color indicates no fluorescence, Red color indicates fluorescence.|width=300px}} | + | {{Team:Aachen/FigureFloatRight|Aachen_K131026_Platereader.gif|title=Testing K131026 in our sensor chips|subtitle=K131026 in our sensorchip induced with 0.2 µl 3-oxo-C{{sub|12}} HSL and measured with a Platereader. Blue color indicates no fluorescence, Red color indicates fluorescence.|width=300px}} |
- | As a next step we used [http://parts.igem.org/Part:BBa_K131026 K131026] from the 2008 iGEM Team of Calgary in our sensor chips to detect 3-oxo-C{{sub|12}} HSL which is produced by ''Pseudomonas aeruginosa''. Firstly we tested it with direct induction of purified 3-oxo-C{{sub|12}} HSL (0 | + | As a next step we used [http://parts.igem.org/Part:BBa_K131026 K131026] from the 2008 iGEM Team of Calgary in our sensor chips to detect 3-oxo-C{{sub|12}} HSL which is produced by ''Pseudomonas aeruginosa''. Firstly we tested it with direct induction of purified 3-oxo-C{{sub|12}} HSL (0.2 µl, 500 µg/ml). Fluorescence measurement was taken every 15 min with an excitation of 496 nm and an emission of 516 nm (for GFP). |
The measured fluorescence showed again a clear fluorescence signal in the induced chip (bottom) compared with the uninduced chip (top). The fluorescence clearly starts in the middle of the chip (point of induction) and then extends outwards, still showing and ever increasing signal of fluorescence. The still measurable fluorescence, even though a lot lower than the bottom chip, is attributed to general leakiness of the promoter and general background fluorescence of growing ''E. coli'' cells. The difference between the induced and non induced chips indicate a clear response to the HSL and a proof for the ability of our sensor chip design to being able to detect the HSL produced by ''Pseudomonas aeruginosa''. | The measured fluorescence showed again a clear fluorescence signal in the induced chip (bottom) compared with the uninduced chip (top). The fluorescence clearly starts in the middle of the chip (point of induction) and then extends outwards, still showing and ever increasing signal of fluorescence. The still measurable fluorescence, even though a lot lower than the bottom chip, is attributed to general leakiness of the promoter and general background fluorescence of growing ''E. coli'' cells. The difference between the induced and non induced chips indicate a clear response to the HSL and a proof for the ability of our sensor chip design to being able to detect the HSL produced by ''Pseudomonas aeruginosa''. |
Revision as of 12:01, 16 October 2014
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