Team:Lyon-INSA-ENS/Realisation/Week2

From 2011.igem.org

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     <h1 style="color: white;"> Week 2 </h1>     
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     <h1 style="color: white;"> Week 3 </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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   <h6 style="text-align :left"> Monday </h6>
   <h6 style="text-align :left"> Monday </h6>
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One transformed colony 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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   <h6 style="text-align :left"> Tuesday </h6>
   <h6 style="text-align :left"> Tuesday </h6>
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4 more new clones have grown : start of 5mL liquid cultures on these clones.
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  <h6 style="text-align :left"> Wednesday </h6>
  <h6 style="text-align :left"> Wednesday </h6>
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Miniprep on the four clones. Verification of plasmid by digestion (EcoRI, HindIII)
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  <h6 style="text-align :left"> Thursday </h6>
  <h6 style="text-align :left"> Thursday </h6>
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Culture of NM 522 cells for later transformations
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PCR from genomic DNA from E.coli strain MC4100 for three genes : RcnR (cobalt receptor), CsgAB (curli operon, part 1) and CsgEFG (curli operon, part 2)
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  <h6 style="text-align :left"> Friday </h6>
  <h6 style="text-align :left"> Friday </h6>
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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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Revision as of 07:40, 28 July 2011










Week 3


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





Monday

One transformed colony 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

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



Wednesday

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



Thursday

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



Friday