Team:Lyon-INSA-ENS/Realisation/Week9

From 2011.igem.org

(Difference between revisions)
(Created page with "<html> <style type="text/css"> #menu { width: 170px ; - →220: background:white; margin-left: 15px; } #menu ul { padding: 0; width: 150px; bord...")
Line 1: Line 1:
 +
{{Lyon-INSA-ENS/header}}
 +
{{Lyon-INSA-ENS/blocStyle}}
 +
{{INSA-Lyon/styletestaurelie}}
 +
{{Lyon-INSA-ENS/menuhorizontal}}
 +
{{Lyon-INSA-ENS/menuNotebookW9}}
 +
<html>
<html>
-
<style type="text/css">
+
<body>
-
#menu {
+
     <div id="language";>
-
     width: 170px ; /*220*/
+
        <img src="https://static.igem.org/mediawiki/2011/0/0e/Drapeau_francais.jpg"; width=20px; /> <a href="/Team:Lyon-INSA-ENS/Realisation/Week7Fr">Version Francaise</a>
-
    background:white;
+
    </div>
-
    margin-left: 15px;
+
-
    }
+
-
#menu ul {
+
<div class="contenugrand2";>
-
    padding: 0;  
+
-
    width: 150px;
+
-
    border:0px solid;
+
-
    margin:0px;
+
-
    margin-top:100px;
+
-
    }
+
-
#menu ul li {
+
     <br/> <br/>
-
     background: #333333 ;
+
     <br/> <br/>
-
    color:#008000  ;
+
     <br>
-
    position:relative;
+
-
    list-style: none; /*on enleve les icones de liste*/  
+
-
     width: 150; /*200*/
+
-
     border: 10px solid;
+
-
    border-color : #333333 ;
+
-
    -moz-border-radius: 10px;
+
-
    -webkit-border-radius: 10px;
+
-
    -goog-ms-border-radius: 10px;
+
-
    border-radius:10px;
+
-
    }
+
-
#menu ul ul {
+
<div class="cadre" ; style="background-color:green;" >
-
    position: relative;  
+
     <br/>
-
     top: 0;
+
     <h1 style="color: white;"> Week 7 </h1>      
-
     left: 0px; /* correspond au décalage a droite*/
+
</div>
-
     display : none
+
-
    }
+
-
#menu ul ul li {
+
<br/>
-
     background: #32CD32;
+
     <p style="text-align : center"> <small> From Monday the 25th of July to Friday the 29th of July 2011 </small> </p>
-
    position:relative;
+
-
    list-style: none; /*on enleve les icones de liste */
+
-
    border-top: 1px solid;
+
-
    border-bottom:1px solid;
+
-
    text-align: center ;
+
-
    }
+
-
#menu li a {
+
     <br/> <br/>
-
    text-decoration: none ;
+
     <br/> <br/>
-
    color : white;
+
     <br/> <br/>
-
    font-family : Times;
+
-
    font-weight :bold;
+
-
     font-size : large;
+
-
     margin-left : 15px;
+
-
     }
+
-
#menu li.sousmenu {
+
  <h6 style="text-align :left"> Monday </h6> <HR>
-
    background: #008000 ;
+
 
-
    }
+
  <br/>
 +
<p style=" line-height : 1.5em">
 +
Miniprep of some other parts from the transformed bacteria. Digestion and electrophoresis to control the presence of the right insert.<br/> <br/>
-
#menu li:hover {
+
Miniprep of bacteria with mutated part CsgAB. Digestion to check restriction profile (removal of intern PstI site).
-
     background:#228B22;
+
     </p>
-
    border-color:#228B22;
+
-
    }
+
-
#menu ul li#actif {
+
    <br/> <br/>
-
        float: left;
+
-
        height: auto;
+
-
        border: 15px solid #555;
+
-
        border-color :#666666;
+
-
        width: 120px;
+
-
        font-family : Times;
+
-
font-weight: bold;
+
-
        font-size : large;
+
-
color: white;
+
-
text-decoration: none;
+
-
        background:#666666;
+
-
        position : relative;
+
-
}
+
 +
  <h6 style="text-align :left"> Tuesday </h6> <HR>
-
</style>
+
    <br/> <br/>  
 +
<p style=" line-height : 1.5em">
 +
Start of a compared culture of a strain transformed with 18A ( constitutive promoter ) only and 18A+OmpR234 ( part which is supposed to induce the formation of a biofilm ) in LB medium, 37°C to check the qualitative behaviour of the OmpR234 part.<br/>
 +
OmpR234 part seems to make the bacteria adherent, the experiment was restarted with a different protocol ( LB diluted by 2 in water, 30°C ) to enhance the adherence.<br/> <br/>
 +
</p>
 +
<br/>
 +
 +
<div style="border:1px solid black;float:left;margin-left:3%; width : 200px">
 +
    <img src="https://static.igem.org/mediawiki/2011/7/74/Boite-petri.jpg" width="200px"/>   
 +
</div>
 +
 
 +
   
 +
<p style=" line-height : 1.5em; margin-left : 35%">
 +
Start of 5mL liquid cultures for minipreps : 18A-OmpR234, 2M-2L Tet, 2M-2L-Kan, rcn-curli, 2M-24E, NiCoT ( a transporter for cobalt that makes the bacteria import cobalt from the medium) <br/> <br/>
-
<div id="menu">
+
Send of plasmid with the right profile for part CsgAB to sequencing. Extraction of new MC4100 strain's DNA. PCR with fresh DNA and Taq Phusion.
-
  <ul class="niveau1">
+
    </p>   
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Notebook/BeforeStarting"> Before starting</a></li>
+
-
          <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Notebook/Week1">Week 1</a></li>
+
-
          <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week2">Week 2</a></li>
+
-
          <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week3">Week 3</a></li>
+
-
          <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week4">Week 4</a></li>
+
-
          <p>
+
-
              <br/><br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week5">Week 5</a></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week6">Week 6</a></li>   
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week7">Week 7</a></li> 
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week8">Week 8</a></li> 
+
-
          <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li id="actif"><U>Week 9</U></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
              <br/> <br/>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week10">Week 10</a></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week11">Week 11</a></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week12">Week 12</a></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week13">Week 13</a></li>
+
-
            <p>
+
-
              <br/> <br/>
+
-
          </p>
+
-
          <li><a href="/Team:Lyon-INSA-ENS/Realisation/Week14">Week 14</a></li>     
+
-
        </ul>         
+
-
        </ul>
+
-
    </li>
+
-
</div>
+
 +
    <br/> <br/> 
 +
    <br/> <br/> 
 +
 +
 +
<h6 style="text-align :left"> Wednesday </h6> <HR>
 +
 +
    <br/>
 +
 +
    <p style=" line-height : 1.5em">
 +
 +
Midiprep and nanodrop of the following ligated or newly obtained plasmids :<br/>
 +
NiCoT : 112,2 ng/µL <br/>
 +
2M-2L-Tet : 109.6 ng/µL <br/>
 +
2M-2L-Kan : 128 ng/µL <br/>
 +
18A-OmpR234 : 59.5 ng/µL <br/>
 +
rcn-Curli : 116 ng/µL <br/>
 +
2M-24E : 92.3 ng/µL <br/><br/>
 +
 +
Fermentas digestion : 2M-2L-Tet (X+P), 2M-2L-Kan (X+P), rcn-curli(E+P), 2M-24E(X+P), Curli(S+P)
 +
<br/><br/>
 +
 +
PCR did not work.
 +
    </p> 
 +
 +
    <br/> <br/>
 +
    <br/> <br/>
 +
 
 +
 +
<h6 style="text-align :left"> Thursday </h6> <HR>
 +
 +
    <br/>
 +
 +
<p style=" line-height : 1.5em">
 +
Standard ligation of :<br/>
 +
-Prcn into Cn backbone<br/>
 +
-rcn-curli into Cn backbone<br/>
 +
-Pcurli into 2M-2L-Kan <br/>
 +
-Pcurli into 2M-2L-Tet <br/><br/>
 +
 +
TSS transformation into NM522.
 +
 +
    </p> 
 +
 +
    <br/> <br/>
 +
 +
<h6 style="text-align :left"> Friday </h6>
 +
<p style=" line-height : 1.5em">
 +
 +
    </p> 
 +
 +
    <br/> <br/>
 +
    <br/> <br/>
 +
 +
 +
</div>
 +
</body>
</html>
</html>
 +
 +
{{Lyon-INSA-ENS/footer}}

Revision as of 08:51, 16 August 2011









Week 7


From Monday the 25th of July to Friday the 29th of July 2011







Monday

Miniprep of some other parts from the transformed bacteria. Digestion and electrophoresis to control the presence of the right insert.

Miniprep of bacteria with mutated part CsgAB. Digestion to check restriction profile (removal of intern PstI site).



Tuesday


Start of a compared culture of a strain transformed with 18A ( constitutive promoter ) only and 18A+OmpR234 ( part which is supposed to induce the formation of a biofilm ) in LB medium, 37°C to check the qualitative behaviour of the OmpR234 part.
OmpR234 part seems to make the bacteria adherent, the experiment was restarted with a different protocol ( LB diluted by 2 in water, 30°C ) to enhance the adherence.


Start of 5mL liquid cultures for minipreps : 18A-OmpR234, 2M-2L Tet, 2M-2L-Kan, rcn-curli, 2M-24E, NiCoT ( a transporter for cobalt that makes the bacteria import cobalt from the medium)

Send of plasmid with the right profile for part CsgAB to sequencing. Extraction of new MC4100 strain's DNA. PCR with fresh DNA and Taq Phusion.





Wednesday

Midiprep and nanodrop of the following ligated or newly obtained plasmids :
NiCoT : 112,2 ng/µL
2M-2L-Tet : 109.6 ng/µL
2M-2L-Kan : 128 ng/µL
18A-OmpR234 : 59.5 ng/µL
rcn-Curli : 116 ng/µL
2M-24E : 92.3 ng/µL

Fermentas digestion : 2M-2L-Tet (X+P), 2M-2L-Kan (X+P), rcn-curli(E+P), 2M-24E(X+P), Curli(S+P)

PCR did not work.





Thursday

Standard ligation of :
-Prcn into Cn backbone
-rcn-curli into Cn backbone
-Pcurli into 2M-2L-Kan
-Pcurli into 2M-2L-Tet

TSS transformation into NM522.



Friday








ENS assystem Biomérieux INSA INSA
Retrieved from "http://2011.igem.org/Team:Lyon-INSA-ENS/Realisation/Week9"