Team:Evry/Interlab Study/Used Devices

From 2014.igem.org

(Difference between revisions)
Line 35: Line 35:
   <center>Plasmid map 2: BBa_J61002 plasmid map with restriction sites ( EcoRI, PstI, SpeI and XbaI)</center>
   <center>Plasmid map 2: BBa_J61002 plasmid map with restriction sites ( EcoRI, PstI, SpeI and XbaI)</center>
   </div>
   </div>
-
 
+
</div>
<br>
<br>
<div class="thumb tnone" style="width:50%;heigth:50%;"/>
<div class="thumb tnone" style="width:50%;heigth:50%;"/>
Line 45: Line 45:
</div>
</div>
</div>
</div>
 +
  </div>
 +
</div>
<h2>Construction way</h2>
<h2>Construction way</h2>
-
<div class="container">
+
<br>
-
     <div class="col-md-6"><img alt="IMAGE" src="https://static.igem.org/mediawiki/2014/a/a6/Schema_interlab_julie.png" width="512px" class="thumbimage"/><br/><center/><br/>
+
     <center><img alt="IMAGE" src="https://static.igem.org/mediawiki/2014/a/a6/Schema_interlab_julie.png" width="512px" class="thumbimage"/><br/><center/><br/>
-
</div>
+
-
<div class="col-md-6" align="justify">
+
<div align="justify">
     To test the expression of promoters, they are cloned into the PSB1C3 vector followed by the  
     To test the expression of promoters, they are cloned into the PSB1C3 vector followed by the  
     <a id="igem-link" target="_blank" href="http://parts.igem.org/Part:BBa_E0240" title="Go to iGEM site">BBa_E0240</a>, a GFP generator. Constructions are inserted in E. coli DH5 alpha that grown on LB medium added with the selection agent, chloramphenicol.
     <a id="igem-link" target="_blank" href="http://parts.igem.org/Part:BBa_E0240" title="Go to iGEM site">BBa_E0240</a>, a GFP generator. Constructions are inserted in E. coli DH5 alpha that grown on LB medium added with the selection agent, chloramphenicol.
   </div>
   </div>
-
</div>
+
<h2>Analysis protocol</h2>
-
<h2>Analysis protocol</h2>
 
-
<br>
 
<div align="justify">
<div align="justify">
  Our team decided to test the fluorescence with a   
  Our team decided to test the fluorescence with a   
Line 74: Line 73:
   Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.<br>
   Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.<br>
   </div>
   </div>
-
</div>
 
</html>
</html>
{{:Team:Evry/Template:HomeFooter}}
{{:Team:Evry/Template:HomeFooter}}

Revision as of 16:55, 17 October 2014

IGEM Evry 2014

Interlab study - Used devices

Used Biobricks and plasmids

IMAGE
Promoter name, plate and well location into the iGEM distribution kit 2014, vector, resistance cassette, part size in bp, used primers and PCR product profile.

IMAGE
Plasmid map 1: PSB1C3 map with restriction sites (NotI, BsaI, EcoRI, PstI, SpeI and XbaI)

IMAGE
Plasmid map 2: BBa_J61002 plasmid map with restriction sites ( EcoRI, PstI, SpeI and XbaI)

IMAGE
Plasmid map 3: BBa_J61002 plasmid map with restriction sites and J23101 promoter ( EcoRI, PstI, SpeI and XbaI)

Construction way


IMAGE

To test the expression of promoters, they are cloned into the PSB1C3 vector followed by the BBa_E0240, a GFP generator. Constructions are inserted in E. coli DH5 alpha that grown on LB medium added with the selection agent, chloramphenicol.

Analysis protocol

Our team decided to test the fluorescence with a TECAN infiniteM200 in 96 well plates.
Fluorescence data were analyzed via following formula that come from 2010 De Jong et al Experimental and computational validation of models of fluorescent and luminescent reporter genes in bacteria .

IMAGE

Equation A: A(t) is the corrected absorbance of DH5 alpha with the functional reporter system, Au(t)the uncorrected absorbance of DH5 alpha with the functional reporter system and Ab(t)the background absorbance corresponding to LB with chloramphenicol medium absorbance.
Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.