Difference between revisions of "Team:Evry/Project/Biosensor"

Line 37: Line 37:
 
<p class="text-justify">Cloning steps were successful as showed sequencing data and colony PCR (figure 2). To remove the promoter GAL1 from our plasmid pYGG1 and replace it with the HRE/CMV promoter, we performed site-directed mutagenesis. First, we amplified the plasmid around GAL1 to remove it. Then, we phospholyrated with a kinase the blunt ends, we added DpnI to remove the template DNA and we ligated with T4 kinase. Colony PCR followed by digestion confirmed the successful assembly by golden gate:</p>
 
<p class="text-justify">Cloning steps were successful as showed sequencing data and colony PCR (figure 2). To remove the promoter GAL1 from our plasmid pYGG1 and replace it with the HRE/CMV promoter, we performed site-directed mutagenesis. First, we amplified the plasmid around GAL1 to remove it. Then, we phospholyrated with a kinase the blunt ends, we added DpnI to remove the template DNA and we ligated with T4 kinase. Colony PCR followed by digestion confirmed the successful assembly by golden gate:</p>
  
<img border="0" class='img-responsive' width="500" src="https://static.igem.org/mediawiki/2015/7/7e/Manquante2.png" alt="" />
+
<img border="0" class='img-responsive' width="500" src="https://static.igem.org/mediawiki/2015/a/a2/Gel_jpg.jpg " alt="" />
 
<p class="text-justify"><strong> Figure 2: Colony PCR for biosensor 2 (HIF alpha and beta) and for biosensor 3 (HRE-CMV-RFP)</strong></p>
 
<p class="text-justify"><strong> Figure 2: Colony PCR for biosensor 2 (HIF alpha and beta) and for biosensor 3 (HRE-CMV-RFP)</strong></p>
  

Revision as of 00:13, 19 September 2015

Scroll to top To top