Team:Paris/August 19

=Screening of the cloning of OmpR*, EnvZ* and FlhDC+promotor=

Minipreps and glycerol stock

 * The clones L133.2 and L133.3 didn't grow up in LB+ampicillin: they seem not to have the plasmid, as revealed by PCR.
 * Minipreps and Glycerol stocks were made for the clones L133.1 and L134.2.


 * ==>Minipreps of L133.1 and L134.2 will be sequenced.

=Screening of the cloning of E0240 and FlhDC+promotor=

Spreading the clones in order to obtain single colonies
The PCR screening of the transformants L139 and L142 of august 15th revealed several bands for a given clone including one band appearing at the right size. There are 2 hypothesis: In order to check these 2 hypothesis and to isolate (if it is possible) the right clone (containing the plasmid with the insert). We decided to spread the "clone" in question in a LB plate in order to carry out a PCR screening on single colonies.
 * The right clone was contaminated by a wrong one
 * The clone contains 2 plasmids: one with the insert and another one without the insert
 * Take of some bacteria from the glycerol stock
 * Resuspension in 400 µL of LB+antibiotic
 * Spreading of 200 µL in a LB agar plate containing the appropriate antibiotic
 * Incubation overnight at 37°C

=Promoter characterization plasmids=

Ligation
Protocol

Transformation
Protocol

These transformations were made during the day at 16°C

Measurement of concentration of minipreps
to be modified standard protocol

Digestion
Protocol Digestion

We had a problem with a gel and we lost these digestions.