Team:Northwestern/Notebook/Protocols/Antibiotic Stocks

From 2011.igem.org

RETURN TO IGEM 2010


Ampicillin



Stocks & Usage

  • Stock Concentration - 50mg/ml in H2O
  • Aliquots - 100μl and 500μl
  • Working Concentration - 50μg/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.


Kanamycin



Stocks & Usage

  • Stock Concentration - 10mg/ml in H2O
  • Aliquots - 200μl and 1ml
  • Working Concentration - 20μg/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.


Tetracycline



Stocks & Usage

  • Stock Concentration - 5mg/ml in 70% Ethanol
  • Aliquots - 200μl and 1ml
  • Working Concentration - 20μg/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



Stocks & Usage

  • Stock concentration - 34mg/ml in 100% Ethanol
  • Aliquots - 1ml
  • Working concentration = 25μ/ml (Stringent), 170μ/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.


Adapted from OpenWetWare