Endy:Preparing Antibiotic Stocks: Difference between revisions
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*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 == | == [[Chloramphenicol]] == | ||
* '''Stock concentration''' - 34mg/ml in | * '''Stock concentration''' - 34mg/ml in 100% Ethanol | ||
* '''Aliquots''' - 1ml | * '''Aliquots''' - 1ml | ||
* '''Working concentration''' = 25μ/ml (Stringent), 170μ/ml (relaxed) | * '''Working concentration''' = 25μ/ml (Stringent), 170μ/ml (relaxed) | ||
| Line 63: | Line 63: | ||
** N.B. There is no need to filter sterilize, as it is in EtOH. | ** N.B. There is no need to filter sterilize, as it is in EtOH. | ||
*Store at -20C. | *Store at -20C. | ||
==Streptomycin== | ==Streptomycin== | ||
| Line 70: | Line 69: | ||
*'''Aliquots''' - 0.5 ml | *'''Aliquots''' - 0.5 ml | ||
*'''Working Concentration''' - 10-50μg/ml | *'''Working Concentration''' - 10-50μg/ml | ||
===Preparation of | ===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). | *Streptomycin is kept in the 4C fridge in 68-564D. I don't think it is light sensitive (not stored in opaque bottle). | ||
*Weigh | *Weigh 2g of streptomycin into a small weigh boat. | ||
*Add | *Add 40ml of milliQ to a 100 or 250ml bottle. | ||
*Add the streptomycin to the milliQ | *Add the streptomycin to the milliQ | ||
*Mix/vortex so all the streptomycin goes into solution. | *Mix/vortex so all the streptomycin goes into solution. | ||
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.
