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Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-16-2014
From 2014.igem.org
(Difference between revisions)
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| - | <u>Tests of antibiotics' stocks</u> <br> | + | <b><u>Tests of antibiotics' stocks</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> | ||
| + | <br> | ||
<ul> | <ul> | ||
<li> Chloramphénicol | <li> Chloramphénicol | ||
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<li> Tetracyclin | <li> Tetracyclin | ||
</ul> | </ul> | ||
| + | <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>Survivability tests</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>Pre-cultures</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> | ||
<u>From glycerol stocks:</u><br> | <u>From glycerol stocks:</u><br> | ||
| + | <br> | ||
<ul> | <ul> | ||
<li>Bl21 | <li>Bl21 | ||
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<li>DH5a pyr tranformed with pMK2 (KanR) | <li>DH5a pyr tranformed with pMK2 (KanR) | ||
</ul> | </ul> | ||
| + | <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. | ||
Revision as of 15:17, 4 September 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
- DH5a tranformed with pCB1C3 (CamR)
- Top10 transformed with pQexp (ErmR)
- DH5a pyr tranformed with pMK2 (KanR)
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
