Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-16-2014

Tests of antibiotics' stocks Six plates of LB agar were made. Five of them contained one of those antibiotics in the dilution 1:1000:

• Chloramphénicol
• Kanamycin
• Erythromycin
• Ampicilin
• Tetracyclin

(The last one was the control of the growth of our bacteria without antibiotics) We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C. Survivability tests Two plates of MB 1X and M9 1X were made and divised in two parts. Each plate was sowed with 5µL of a liquid MB 1X culture of Pseudovibrio denitrificans(dated 12th August) in a part and with 5µL of a liquid M9 1X culture Pseudovibrio denitrificans (dated 12th August) in the other part. Pre-cultures Cultures in MB 1X and M9 1X of Pseudovibrio were passed by a 1/10 dilution and let incubate overnight at 30°C. New cultures of different E.Coli were also started from glycerol or plate and let incubated overnight at 37°C. From glycerol stocks:

• Bl21
• Top10
• DH5a

From plates:

• DH5a tranformed with pCB1C3
• Top10 transformed with pQexp
• DH5a pyr tranformed with pMK2

Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection. For each medium we make a negative contrôle without bacteria.

Aug 16