Fong:Antibiotics
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Protocols for the preparation of antibiotic stock solutions
Ampicillin (Amp)
- Stock concentration = 100mg/mL
- Stock preparation protocol:
- Weigh out 1.0g of Amp and place in dry 50mL conical tube
- Add 10mL of pure water and dissolve the Amp
- Remove the solution using a sterile syringe
- Filter the solution using a syringe and filter pad (as it exits) into another sterile 50mL tube
- Aliquot 1.0mL samples into 10 sterile microfuge tubes
- Label and store in -20C freezer
- Working concentration = 100μg/mL (add 1mL of stock per liter of media)
Kanamycin (Kan)
- Stock concentration = 30mg/mL
- Stock preparation protocol:
- Weigh out 300mg of Kan and place in dry 50mL conical tube
- Add 10mL of pure water and dissolve the Kan
- Remove the solution using a sterile syringe
- Filter the solution (using a filter at the end of the syringe) and deposit into a sterile 15mL conical tube
- Split solution into ten sterile microfuge tubes (1mL aliquots)
- Label and store in -20C freezer
- Working concentration = 30μg/mL (add 1mL of stock per liter of media)
Chloramphenicol (Cm)
- Stock concentration = 25mg/mL
- Stock preparation protocol:
- Weigh out 500mg of Cm and place in dry 50mL conical tube
- Add 20mL of 100% ethanol and dissolve Cm by vortexing
- There is no need to filter since Cm is stored in 100% ethanol
- Aliquot 1.0mL samples into 20 sterile microfuge tubes
- Label and store in -20C freezer
- Working concentration = 25μg/mL (add 1mL of stock per liter of media)
Tetracycline (Tet)
- Stock concentration = 15mg/mL
- Stock preparation protocol:
- Weigh out 300mg of Tet and place in dry 50mL conical tube
- Add 20mL of 70% ethanol and dissolve Tet by vortexing
- Remove the solution using a sterile syringe
- Filter the solution (using a filter at the end of the syringe) and deposit into a sterile 50mL conical tube
- Aliquot 1.0mL samples into 20 sterile microfuge tubes
- Label and store in -20C freezer
- Working concentration = 15μg/mL (add 1mL of stock per liter of media)