Fong:Antibiotics

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)