Team:Cambridge/Protocols/AntibioticStocks

Use of Antibiotic Stocks

Risk Assessment

Stocks are to be kept in a -20^oc 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.