Endy:Preparing Antibiotic Stocks

Stocks & Usage

 * Stock Concentration - 50mg/ml in H2O
 * Aliquots - 100&mu;l and 500&mu;l
 * Working Concentration - 50&mu;g/ml

Preparation of 80ml stock solution

 * Ampicillin is kept in the 4C fridge in 68-564D. It is light sensitive.
 * Weigh 4g of ampicillin into a small weigh boat.
 * Add 80ml of milliQ to a 250ml bottle.
 * Add the ampicillin to the milliQ
 * Mix/vortex so all the ampicillin goes into solution.
 * Filter sterilize the solution into a falcon tube using a 20ml syringe and a 200nm filter.
 * Aliquot into pcr tubes and 1.7ml eppendorfs.
 * Store at -20C.
 * Store the small aliquots in the small box and the big aliquots in the larger box.

Stocks & Usage

 * Stock Concentration - 10mg/ml in H2O
 * Aliquots - 200&mu;l and 1ml
 * Working Concentration - 20&mu;g/ml

Preparation of 80ml stock solution

 * Kanamycin is kept in the 4C fridge in 68-564D. It is light sensitive.
 * Weigh 800mg of kanamycin into a small weigh boat.
 * Add 80ml of milliQ to a 250ml bottle.
 * Add the kanamycin to the milliQ
 * Mix/vortex so all the kanamycin goes into solution.
 * Filter sterilize the solution into a falcon tube using a 20ml syringe and a 200nm filter.
 * Aliquot into pcr tubes and 1.7ml eppendorfs.
 * Store at -20C.
 * Store the small aliquots in the small box and the big aliquots in the larger box.

Stocks & Usage

 * Stock Concentration - 5mg/ml in 70% Ethanol
 * Aliquots - 200&mu;l and 1ml
 * Working Concentration - 20&mu;g/ml

Preparation of 80ml stock solution

 * Tetracycline is kept in the 4C fridge in 68-564D. It is light sensitive.
 * Weigh 400mg of tetracyline HCL into a small weigh boat.
 * Dilute 95% Ethanol to 70% using milliQ water.
 * Adding 20ml of milliQ to 60ml of 95% ethanol gives 80ml of 71% ethanol.
 * Add 80ml of 70% Ethanol to a 250ml bottle.
 * Add the tetracycline HCL to the ethanol.
 * Mix/vortex vigorously so all the tetracycline goes into solution.
 * Filter sterilize the solution into a falcon tube using a 20ml syringe and a 200nm filter.
 * Aliquot into pcr tubes and 1.7ml eppendorfs.
 * Store at -20C and protect any unused stock solution from light.
 * Store the small aliquots in the small box and the big aliquots in the larger box.

Chloramphenicol

 * Stock concentration - 34mg/ml in 100% Ethanol
 * Aliquots - 1ml
 * Working concentration = 25&mu;/ml (Stringent), 170&mu;/ml (relaxed)

Preparation of 80ml stock solution

 * Chloramphenicol is kept at room temperature. It is near our other chemicals
 * Weight 2.72g of chloramphenicol sulfate into a small weight boat.
 * Add 80ml 100% EtOH to the chloramphenicol
 * 95% EtOH probably works just fine, but I haven't tried it.
 * Mix/vortex vigorously so all the chloramphenicol goes into solution.
 * Aliquot into 1.7ml eppendorfs.
 * N.B. There is no need to filter sterilize, as it is in EtOH.
 * Store at -20C.

Stocks & Usage

 * Stock Concentration: 50mg/ml in H2O
 * Aliquots - 0.5 ml
 * Working Concentration - 10-50&mu;g/ml

Preparation of 40ml stock solution

 * Streptomycin is kept in the 4C fridge in 68-564D. I don't think it is light sensitive (not stored in opaque bottle).
 * Weigh 2g of streptomycin into a small weigh boat.
 * Add 40ml of milliQ to a 100 or 250ml bottle.
 * Add the streptomycin to the milliQ
 * Mix/vortex so all the streptomycin goes into solution.
 * Filter sterilize the solution into a falcon tube using a 60ml syringe and a 200nm filter.
 * Aliquot into 1.7ml eppendorfs.
 * Store at -20C.