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Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-16-2014
From 2014.igem.org
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| - | <u>Tests of antibiotics' stocks</u> <br> | + | <b><u><big><FONT COLOR=#003333>Tests of antibiotics' stocks</font></big></u></b> <br> |
Six plates of LB agar were made. | Six plates of LB agar were made. | ||
Five of them contained one of those antibiotics in the dilution 1:1000:<br> | Five of them contained one of those antibiotics in the dilution 1:1000:<br> | ||
| - | + | ||
<li> Chloramphénicol | <li> Chloramphénicol | ||
<li> Kanamycin | <li> Kanamycin | ||
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<li> Ampicilin | <li> Ampicilin | ||
<li> Tetracyclin | <li> Tetracyclin | ||
| - | < | + | |
| + | <br> | ||
(The last one was the control of the growth of our bacteria without antibiotics) | (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.<br> | We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C.<br> | ||
| + | <br> | ||
| - | <u>Survivability tests</u><br> | + | <b><u><big><FONT COLOR=#003333>Survivability tests</font></big></u></b><br> |
Two plates of MB 1X and M9 1X were made and divised in two parts. | 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.<br> | 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.<br> | ||
| + | <br> | ||
| - | + | <u><b><big><FONT COLOR=#003333>Pre-cultures</font></big></u></b> <br> | |
Cultures in MB 1X and M9 1X of Pseudovibrio were passed by a 1/10 dilution and let incubate overnight at 30°C. | 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.<br> | New cultures of different E.Coli were also started from glycerol or plate and let incubated overnight at 37°C.<br> | ||
| - | < | + | <br> |
| - | + | ||
| + | From glycerol stocks:<br> | ||
<li>Bl21 | <li>Bl21 | ||
<li>Top10 | <li>Top10 | ||
<li>DH5a | <li>DH5a | ||
| - | < | + | <br> |
| - | < | + | <br> |
| - | + | From plates: | |
<li>DH5a tranformed with pCB1C3 (CamR) | <li>DH5a tranformed with pCB1C3 (CamR) | ||
<li>Top10 transformed with pQexp (ErmR) | <li>Top10 transformed with pQexp (ErmR) | ||
<li>DH5a pyr tranformed with pMK2 (KanR) | <li>DH5a pyr tranformed with pMK2 (KanR) | ||
| - | < | + | <br> |
| + | <br> | ||
Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection. | Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection. | ||
For each medium we make a negative contrôle without bacteria. | For each medium we make a negative contrôle without bacteria. | ||
Latest revision as of 15:21, 12 October 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:
(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:
From plates:
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
