Difference between revisions of "Team:SCSU-New Haven/Experiments"

 
Line 53: Line 53:
 
<hr/>
 
<hr/>
 
<p class="header_intro">  The Degrader would then be added and measurements of Indigo Dye would be taken. Comparing these results to the production of indole would tell us how well the Degrader enzymes convert indole to indigo dye in the system. We could also assay the conversion of 1-MethylNapthalene to 3-Methylsalicylate by comparing the fluorescence produced when 1-MethylNapthalene is added compared to 3-Methylsalicylate.   
 
<p class="header_intro">  The Degrader would then be added and measurements of Indigo Dye would be taken. Comparing these results to the production of indole would tell us how well the Degrader enzymes convert indole to indigo dye in the system. We could also assay the conversion of 1-MethylNapthalene to 3-Methylsalicylate by comparing the fluorescence produced when 1-MethylNapthalene is added compared to 3-Methylsalicylate.   
</p>
+
</p></div>
<img src="https://static.igem.org/mediawiki/2016/c/c2/SCSU-New_HavenDegraderAssay.jpeg">  
+
<center><img src="https://static.igem.org/mediawiki/2016/c/c2/SCSU-New_HavenDegraderAssay.jpeg"> </center>
 
 
 
</body>
 
</body>
 
</html>
 
</html>
 
{{SCSU-New_Haven/Footer}}
 
{{SCSU-New_Haven/Footer}}

Latest revision as of 23:19, 30 November 2016

SCSU IGEM SCSU IGEM

SCSU iGEM

Experiments

You have the parts. You do the tests to get the numbers.

Detector Assay


The Detector assay will be done by placing GFP downstream of Psal and exposing the cells to 3-Methylsalicylate. The fluorescence response will be measured. We would like to see a response from 0.1uM and less of 3-Methylsalicylate as this was the amount required to produce a response in the paper we obtained the sequence from.

Feedback Assay


The Assay for the Feedback part will be similar to the Detector Assay. The Feedback part will be place downstream of Psal and upstream of GFP and fluorescence will be measured. If the part is working as intended, we would see a rise in fluorescence to a steady state. This means the feedback loop is turning the Psal promoter on and keeping it on as intended.

Color Reporter Assays


The Assay for color reporter will be as the above assays substituting the GFP for the Color Reporter part. The production of Indole will be measured using a spectrophotometer. We should expect to see production of Indole comparable to fluorescence.

Degrader Assay


The Degrader would then be added and measurements of Indigo Dye would be taken. Comparing these results to the production of indole would tell us how well the Degrader enzymes convert indole to indigo dye in the system. We could also assay the conversion of 1-MethylNapthalene to 3-Methylsalicylate by comparing the fluorescence produced when 1-MethylNapthalene is added compared to 3-Methylsalicylate.