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| | <div class="cadre" ; style="background-color:green;" > | | <div class="cadre" ; style="background-color:green;" > |
| | <br/> | | <br/> |
| - | <h1 style="color: white;"> Week 7 </h1> | + | <h1 style="color: white;"> Week 9 </h1> |
| | </div> | | </div> |
| | | | |
| | <br/> | | <br/> |
| - | <p style="text-align : center"> <small> From Monday the 25th of July to Friday the 29th of July 2011 </small> </p> | + | <p style="text-align : center"> <small> From Monday the 15th of August to Friday the 19th of August 2011 </small> </p> |
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| | <br/> <br/> | | <br/> <br/> |
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| | <p style=" line-height : 1.5em"> | | <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/>
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| | | | |
| - | Miniprep of bacteria with mutated part CsgAB. Digestion to check restriction profile (removal of intern PstI site).
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| | </p> | | </p> |
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| | <p style=" line-height : 1.5em"> | | <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/>
| + | </p> |
| - | 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/>
| + | |
| - | | + | |
| - | 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.
| + | |
| - | </p>
| + | |
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| | <br/> <br/> | | <br/> <br/> |
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| | <p style=" line-height : 1.5em"> | | <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/>
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| - | 2M-2L-Kan : 128 ng/µL <br/>
| |
| - | 18A-OmpR234 : 59.5 ng/µL <br/>
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| - | rcn-Curli : 116 ng/µL <br/>
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| - | 2M-24E : 92.3 ng/µL <br/><br/>
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| - |
| |
| - | Fermentas digestion : 2M-2L-Tet (X+P), 2M-2L-Kan (X+P), rcn-curli(E+P), 2M-24E(X+P), Curli(S+P)
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| - | <br/><br/>
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| - |
| |
| - | PCR did not work.
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| | </p> | | </p> |
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| | <p style=" line-height : 1.5em"> | | <p style=" line-height : 1.5em"> |
| - | Standard ligation of :<br/>
| |
| - | -Prcn into Cn backbone<br/>
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| - | -rcn-curli into Cn backbone<br/>
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| - | -Pcurli into 2M-2L-Kan <br/>
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| - | -Pcurli into 2M-2L-Tet <br/><br/>
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| - |
| |
| - | TSS transformation into NM522.
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| - |
| |
| | </p> | | </p> |
| | | | |
"
