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Team:Cambridge/Protocols/AntibioticStocks
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==Use of Antibiotic Stocks== | ==Use of Antibiotic Stocks== | ||
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| + | [[Team:Cambridge/RiskAssessments/AntibioticStocks| Risk Assessment]] | ||
Stocks are to be kept in a -20<sup>o</sup>c freezer, clearly labelled. | Stocks are to be kept in a -20<sup>o</sup>c freezer, clearly labelled. | ||
Revision as of 14:48, 23 September 2012
Contents |
Use of Antibiotic Stocks
Stocks are to be kept in a -20oc freezer, clearly labelled.
Available stocks in the lab are:
- Ampicillin 100mg/ml , 1ml in each eppendorf
- Chloramphenicol 25mg/ml , 200µl in each eppendorf
- Kanamycin 50mg/ml , 1ml in each eppendorf
- Erythromycin 10mg/ml , 1ml in each eppendorf
To make plates:
Put one entire tube into 1 litre of LB AGAR - This should be done in the flow hood.
This will make the standard plates
- Ampicillin 100µg/ml
- Chloramphenicol 5µg/ml
- Kanamycin 50µg/ml
- Erythromycin 5µg/ml
To make these concentrations in the standard 250ml bottle LB Agar, add:
- 250µl ampicillin stock
- 50µl chloramphenicol stock
- 250µl kanamycin stock
- 125µl erythromycin stock
Common plate concentrations for backbones:
- pSB1C3 (E.coli) : 25µg/ml Chloramphenicol
- pSJ150 (E.coli) : 100µg/ml Ampicillin
- pSJ150 (B.subtilis) : 5µg/ml Chloramphenicol
- pSB1K3 (E.coli) : 50µg/ml Kanamycin
- PSB4K5 (E.coli) : 50µg/ml Kanamycin
To make liquid cultures
Check for specific concentrations, this is usually lower than the ones you use for plates, and add the required amount to sterile LB.
