Team:Lyon-INSA-ENS/Realisation/Week9
From 2011.igem.org
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Start of 24 well plate adherence test in M63G medium with PHL818 ( positive control ), NM522 ( negative control ), S4 + Amp + CoCl2 ( in increasing concentration ) <br/> <br/> | Start of 24 well plate adherence test in M63G medium with PHL818 ( positive control ), NM522 ( negative control ), S4 + Amp + CoCl2 ( in increasing concentration ) <br/> <br/> | ||
- | 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 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) <br/><br/> |
+ | |||
Transformation and plating of S19 with : p157(3µL), p115(3µL), 3 replica each. | Transformation and plating of S19 with : p157(3µL), p115(3µL), 3 replica each. | ||
Revision as of 11:41, 18 August 2011
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