Team:Lyon-INSA-ENS/Realisation/Week9

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Week 9


From Monday the 15th of August to Friday the 19th of August 2011







Monday




Tuesday



Start of 5mL cultures for adherence tests from the collection :
LB : NM522
LB/2 : NM522, S3, S4, S15, S19
M63 : NM522, S15, S3, S19, S4, S18

Plating of individual or non individual clones from the previous NM522-pIG16 and NM522-pIG3 cultures on LB+amp medium. Incubation at 37°C overnight.

Preparation of 2X50mL LB/2 solutions.





Wednesday


Start of 3x5mL cultures of NM522 in LB medium from 50µL of a saturated NM522 culture for a later transformation.
Start of 5mL cultures of the individual clones plated on the previous day.

Test of antibiotics : start of 5mL cultures ( 5mL LB, 50µL antibiotic, 10µL saturated NM522) of NM522 with antibiotics to test if the antibiotic is still good for Kan, Tet, Cm and Amp
The bacteria with Amp grow : a new solution is made ( 5x1mL mother solutions at 100mg/mL from solid Amp, and 2x10mL solutions at 100µg/mL by dilution of 10µL of mother solution into 10mL sterile water )

Start of 24 well plate adherence test in M63G medium with PHL818 ( positive control ), NM522 ( negative control ), S4 + Amp + CoCl2 ( in increasing concentration )

Transformation and plating of NM522 with : pIG6 (3µL), pIG7 (3µL), pIG24 (2µL), p56 (2µL), p10 (2µL), pIG16+p157 (3µL each), pIG16+p115(3µL each), pIG16+p127(3µL each), pIG16+p116 ( 3µL and 5µL respectively ), pIG25 (1µL) Transformation and plating of S19 with : p157(3µL), p115(3µL), 3 replica each.





Thursday




Friday








ENS assystem Biomérieux INSA INSA