Endy:Preparing Antibiotic Stocks

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Current revision (13:08, 27 October 2008) (view source)
(→Preparation of 80ml stock solution)
 
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===Stocks & Usage===
===Stocks & Usage===
*'''Stock Concentration''' - 50mg/ml in H<sub>2</sub>O
*'''Stock Concentration''' - 50mg/ml in H<sub>2</sub>O
-
*'''Aliquots''' - 100&mu;l and 100&mu;l
+
*'''Aliquots''' - 100&mu;l and 500&mu;l
*'''Working Concentration''' - 50&mu;g/ml
*'''Working Concentration''' - 50&mu;g/ml
 +
===Preparation of 80ml stock solution===
===Preparation of 80ml stock solution===
-
*Tetracycline is kept in the 4C fridge in 68-564D.  It is light sensitive.
+
*Ampicillin is kept in the 4C fridge in 68-564D.  It is light sensitive.
-
*Weigh 400mg of tetracyline HCL into a small weigh boat.
+
*Weigh 4g of ampicillin into a small weigh boat.
-
*Dilute 95% Ethanol to 70% using milliQ water.
+
*Add 80ml of milliQ to a 250ml bottle.
-
**Adding 20ml of milliQ to 60ml of 95% ethanol gives 80ml of 71% ethanol.
+
*Add the ampicillin to the milliQ
-
*Add 80ml of 70% Ethanol to a 250ml bottle.
+
*Mix/vortex so all the ampicillin goes into solution.
-
*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.
*Filter sterilize the solution into a falcon tube using a 20ml syringe and a 200nm filter.
*Aliquot into pcr tubes and 1.7ml eppendorfs.
*Aliquot into pcr tubes and 1.7ml eppendorfs.
-
*Store at -20C and protect any unused stock solution from light.
+
*Store at -20C.
*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.
==[[Kanamycin]]==
==[[Kanamycin]]==
 +
===Stocks & Usage===
 +
*'''Stock Concentration''' - 10mg/ml in H<sub>2</sub>O
 +
*'''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.
==[[Tetracycline]]==
==[[Tetracycline]]==
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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.
 +
 +
== [[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.
 +
 +
==Streptomycin==
 +
===Stocks & Usage===
 +
*'''Stock Concentration:''' 50mg/ml in H<sub>2</sub>O
 +
*'''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.

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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