Team:Lyon-INSA-ENS/Realisation/Week2

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     <h1 style="color: white;"> Week 2 </h1>     
     <h1 style="color: white;"> Week 2 </h1>     
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     <p style="text-align : center"> <small> From Monday the 13th of June to Friday the 17th of June 2011 </small> </p>
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     <p style="text-align : center"> <small> From Monday the 20th of June to Friday the 24th of June 2011 </small> </p>
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<FONT COLOR="orange"> <b> Extraction of NiCoT </b></FONT> <br><br>
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One transformed colony (NicoT in NM522) seemed correct, the others were probably satellite colonies without the plasmid.<br/>
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Start of 5mL liquid culture of the main colony. <br/>
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Plating of the satellite colonies to check the presence of the plasmid <br/>
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<FONT COLOR="orange"> <b> Extraction of NiCoT </b></FONT> <br><br>
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4 more new clones have grown : start of 5mL liquid cultures on these clones.
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<FONT COLOR="orange"> <b> Extraction of NiCoT </b></FONT> <br><br>
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Miniprep on the four clones. Verification of plasmid by <b>digestion</b> (EcoRI, HindIII)
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Culture of NM 522 cells for later transformations
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<FONT COLOR="green"> <b>PCR and mutagenesis of rcn, csgBA, csgEFG</b> </FONT> <br><br>
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Extraction of plasmid with NicoT gene from filter paper. Transformation of the extracted DNA in NM522 strain using TSS. Culture and selection of transformants on ampicilline plate during the week end.
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<b>PCR</b> from genomic DNA from E.coli strain MC4100 to amplify three genes : RcnR (cobalt receptor), CsgAB (curli operon, part 1) and CsgEFG (curli operon, part 2)
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Latest revision as of 23:34, 21 September 2011







Week 2


From Monday the 20th of June to Friday the 24th of June 2011





Monday

Extraction of NiCoT

One transformed colony (NicoT in NM522) seemed correct, the others were probably satellite colonies without the plasmid.
Start of 5mL liquid culture of the main colony.
Plating of the satellite colonies to check the presence of the plasmid





Tuesday

Extraction of NiCoT

4 more new clones have grown : start of 5mL liquid cultures on these clones.





Wednesday

Extraction of NiCoT

Miniprep on the four clones. Verification of plasmid by digestion (EcoRI, HindIII)





Thursday

PCR and mutagenesis of rcn, csgBA, csgEFG

PCR from genomic DNA from E.coli strain MC4100 to amplify three genes : RcnR (cobalt receptor), CsgAB (curli operon, part 1) and CsgEFG (curli operon, part 2)













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