Fong:Antibiotics

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Protocols for the preparation of antibiotic stock solutions

Contents

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 and place in dry 50mL conical tube
    2. Add 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 and place in dry 50mL conical tube
    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 20 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:
    1. Weigh out 300mg of Tet and place in dry 50mL conical tube
    2. Add 20mL of 70% ethanol and dissolve Tet by vortexing
    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 50mL conical tube
    5. Aliquot 1.0mL samples into 20 sterile microfuge tubes
    6. Label and store in -20C freezer
  • Working concentration = 15μg/mL (add 1mL of stock per liter of media)
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