From Monday the 12th of September to Friday the 16th of September 2011

Monday

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Transformations and controls for future tests

Plasmid and strain Collection

Plating of S17 from the collection to extract a bigger quantity of pIG25 ( pSB1C3 containing rcn-csgBAEFG )

Storage of the correct clone of:
PHL1414 + pIG3 (S30)
PHL1414+pIG16 (S31)
MC4100 + pIG34 (S32)

Storage of rcn-OmpR in psB1K3 (pIG34) from the previous miniprep.

Adherence Test Preparation

Others

Pouring of 9 LB+Amp plates

Dilution of 5µL pIG25 (371.4 ng/µL) into 25µL water to obtain 3 additional tubes at concentration of about 50ng/µL for sequencing.
Sequencing (pIG25 and pIG27) sent.

Tuesday

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Transformations and controls for future tests

Plasmid and strain Collection

Start of a 5mL liquid culture of S17 in LB+Cm medium
Minipreps of this culture to extract pIG25 in larger quantity. Digestion (E+P) and electrophoresis for control : the insert is too small, not correct.

Adherence Test Preparation

Others

Wednesday

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Transformations and controls

The Ptrc strong promoter has been received from Genecust in pBluescriptIISK+: transformation in MC4100 and plating on LB+Amp medium.

Digestion by E+P of this plasmid and ligation into pSB1C3 overnight

Plating of PHL916 on LB ( sureE strain that should not recombine DNA ) in case the transformation of pIG25 into MC4100 fails.

Plasmid and strain Collection

rcn-OmpR234 adherence test

Start of a rcn-OmpR234 24 well plate with :
S27 ( rcn in pSB1C3 ) without cobalt and with cobalt (25µM)
S33 ( rcn-OmpR234 in pSB1C3 ) with increasing concentration of cobalt (0, 10, 25 and 100 µM)

Others

Plating of PHL644, PHL1256, PHL628 to be sent to other research teams, on request.

Thursday

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Friday

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