Endy:Preparing Antibiotic Stocks

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Current revision (12:08, 27 October 2008) (view source)
(Preparation of 80ml stock solution)
 
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*Store at -20C and protect any unused stock solution from light.
*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.
*Store the small aliquots in the small box and the big aliquots in the larger box.
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== [[Chloramphenicol]] ==
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* '''Stock concentration''' - 34mg/ml in 100% Ethanol
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* '''Aliquots''' - 1ml
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* '''Working concentration''' = 25μ/ml (Stringent), 170μ/ml (relaxed)
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=== Preparation of 80ml stock solution ===
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*Chloramphenicol is kept at room temperature. It is near our other chemicals
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*Weight 2.72g of chloramphenicol sulfate into a small weight boat.
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*Add 80ml 100% EtOH to the chloramphenicol
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**95% EtOH probably works just fine, but I haven't tried it.
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*Mix/vortex vigorously so all the chloramphenicol goes into solution.
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*Aliquot into 1.7ml eppendorfs.
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** N.B. There is no need to filter sterilize, as it is in EtOH.
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*Store at -20C.
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==Streptomycin==
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===Stocks & Usage===
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*'''Stock Concentration:''' 50mg/ml in H<sub>2</sub>O
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*'''Aliquots''' - 0.5 ml
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*'''Working Concentration''' - 10-50&mu;g/ml
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===Preparation of 40ml stock solution===
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*Streptomycin is kept in the 4C fridge in 68-564D.  I don't think it is light sensitive (not stored in opaque bottle).
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*Weigh 2g of streptomycin into a small weigh boat.
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*Add 40ml of milliQ to a 100 or 250ml bottle.
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*Add the streptomycin to the milliQ
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*Mix/vortex so all the streptomycin goes into solution.
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*Filter sterilize the solution into a falcon tube using a 60ml syringe and a 200nm filter.
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*Aliquot into 1.7ml eppendorfs.
 +
*Store at -20C.

Current revision

Contents

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

  • 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.

Streptomycin

Stocks & Usage

  • Stock Concentration: 50mg/ml in H2O
  • Aliquots - 0.5 ml
  • Working Concentration - 10-50μg/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.
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