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Team:Evry/Notebook/CellCharacterization/Antibiotic test
From 2014.igem.org
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| - | < | + | <h6>08-16-2014</h6> |
| - | < | + | <h4> <u>Tests of antibiotics' stocks</u> </h4> |
Six plates of LB agar were made. | Six plates of LB agar were made. | ||
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We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C. | We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C. | ||
| - | < | + | <h4> <u>Survivability tests</u> </h4> |
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. | 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. | ||
| - | < | + | <h4> <u>Pre-cultures</u> </h4> |
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. | New cultures of different E.Coli were also started from glycerol or plate and let incubated overnight at 37°C. | ||
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For each medium we make a negative contrôle without bacteria. | For each medium we make a negative contrôle without bacteria. | ||
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Revision as of 09:26, 25 August 2014
Preparation of antibiotic stocks
| Antibiotic | Stock concentration | Protocol |
| Kanamycin | 25 mg/mL | Weight 0.56g of Kanamycin powder, solubilization into 20mL of miliQ water. Vortex 5min and filtration with a 200nm filter. Stock : -20°C (aliquots of 1mL) |
| Streptomycin | 100 mg/mL | Weight 1g of Streptomycin powder, solubilization into 20mL of miliQ water. Vortex 5min and filtration with a 200nm filter. Stock : -20°C (aliquots of 1mL) |
| Amplicilin | 50 mg/mL | Weight 1g of Amplicilin powder, solubilization into 40mL of miliQ water. Vortex 5min and filtration with a 200nm filter. Stock : -20°C (aliquots of 1mL) |
| Tetracyclin | 15 mg/mL | Weight 0.6g of Tetracyclin powder, solubilization into 20mL of ethanol 50%. Vortex 5min and filtration with a 200nm filter. Stock : -20°C and hiden from light(aliquots of 1mL) |
| Chloramphenicol | 34 mg/mL | Weight 0.34g of Chloramphenicol powder, solubilization into 10mL of ethanol 100%. Vortex 5min and filtration with a 200nm filter. Stock : -20°C and hiden from light(aliquots of 1mL) |
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
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
- Top10 transformed with pQexp
- DH5a pyr tranformed with pMK2
