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

Monday

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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

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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

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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-Où234 : 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

Friday