Fong:Antibiotics

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Protocols for the preparation of antibiotic stock solutions

Ampicillin (Amp)

• Stock concentration = 100mg/mL
• Stock preparation protocol:
  1. Weigh out 1.0g of Amp and place in dry 50mL conical tube
  2. Add 10mL of pure water and dissolve the Amp
  3. Remove the solution using a sterile syringe
  4. Filter the solution using a syringe and filter pad (as it exits) into another sterile 50mL tube
  5. Aliquot 1.0mL samples into 10 sterile microfuge tubes
  6. 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:
  1. Weigh out 300mg of Kan
  2. Add Kan to a 50mL conical tube containing 10mL of pure water and dissolve the Kan
  3. Remove the solution using a sterile syringe
  4. Filter the solution (using a filter at the end of the syringe) and deposit into a sterile 15mL conical tube
  5. Split solution into ten sterile microfuge tubes (1mL aliquots)
  6. 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:
  1. Weigh out 500mg of Cm
  2. Add 20mL of 100% ethanol and dissolve Cm by vortexing
  3. There is no need to filter since Cm is stored in 100% ethanol
  4. Aliquot 1.0mL samples into 10 sterile microfuge tubes
  5. 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:
• Working concentration = 15μg/mL (add 1mL of stock per liter of media)