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

Monday

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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

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Extraction of NiCoT

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

Wednesday

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Extraction of NiCoT

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

Thursday

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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)