Team:Cambridge/Protocols/AntibioticStocks

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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.


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