Endy:Preparing Antibiotic Stocks: Difference between revisions
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==Ampicillin== | ==[[Ampicillin]]== | ||
==Kanamycin== | ===Stocks & Usage=== | ||
*'''Stock Concentration''' - 50mg/ml in H<sub>2</sub>O | |||
*'''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 H<sub>2</sub>O | |||
*'''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]]== | ==[[Tetracycline]]== | ||
===Stocks & Usage=== | ===Stocks & Usage=== | ||
*'''Stock Concentration''' - 5mg/ml in 70% Ethanol | *'''Stock Concentration''' - 5mg/ml in 70% Ethanol | ||
*'''Aliquots''' - | *'''Aliquots''' - 200μl and 1ml | ||
*'''Working Concentration''' - 20μg/ml | *'''Working Concentration''' - 20μg/ml | ||
===Preparation of 80ml stock solution=== | ===Preparation of 80ml stock solution=== | ||
Line 16: | Line 47: | ||
*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 | *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''' = 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 H<sub>2</sub>O | |||
*'''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. |
Latest revision as of 10:08, 27 October 2008
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.