From Monday the 5th of September to Friday the 9th of September 2011

Monday

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Transformations and controls for future tests

Methyl violet coloration of the flocculation tests from the previous week.

Start of 3 replica of 24 well plates containing :
MC4100+pIG6, MC4100+pIG6+10µM Co ( controls )
MC4100+pIG2 with increasing concentration in CoCl2 (0, 10, 25, 50, 100 µM )

Revelation of the 2 plates from september 2nd.

Miniprep of clones 1, 2 and 3 of both MC4100+pIG6 and MC4100+pIG2.

Tuesday

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TSS transformation of MC4100 with pIG27 (Cm) or pIG16 (Amp).

CFA tests

Digestion and electrophoresis of the previously extracted DNA of MC4100+pIG6 clones and MC4100+pIG2 clones. Comparison made with the parts put in the collection.

Wednesday

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Results of tuesday transformations : no clone on the negative controls, several on the tests. Isolation of 4 clones per transformation in solid and liquid culture.

Pouring of Amp plates.

Revelation of the 2 plates from september 5th.

Thursday

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E+P digestion and electrophoresis of all the isolated clones for the following transformations : MC4100+pIG27, MC4100 + pIG16, MC4100 + rcn-ompR234.
All clones are correct for pIG27 and pIG16. No clone is correct for rcn-ompR234 -> detection of an error in the ligation.
Storage of clone 1 for pIG27 and pIG16. Plating of those same clones on LB+antibiotic (Cm and Amp respectively ).

Friday

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Nanodrop quantification of the previous pIG27 miniprep : 48.0 ng/µL.
Preparation of tubes for sequencing (30µL at approximately 50-100 ng/µL). The pIG27 miniprep is used as such. pIG25 is diluted from 5µL of the collection (371.4 ng/µL) with 25µL water.