Boston University 2006:making antibiotic stocks
Use the following protocol to produce 5 1mL aliquots of stock solution of your desired antibiotic. Use a 1:1000 dilution to reach a working concentration from the stock (i.e. take 5uL from the Amp stock and add to 5mL of LB broth to produce 100ug/mL Amp + LB broth).
- Powder of given antibiotic
- dH20 and/or 100% ethanol
- 5mL Falcon tube(s)
- 5 1.5mL microcentrifuge tubes (per 5mL falcon tube)
|Ampicillin||Amp||100 ug/mL||100 mg/mL||orange|
|Chloramphenicol||Chm||30 ug/mL||30 mg/mL||in 100% ethanol||green|
|Kanamycin||Kan||50 ug/mL||50 mg/mL||red|
|Tetracycline HCL||Tet||50 ug/mL||50 mg/mL||in 100% ethanol||yellow|
These directions are not correct yet. Antibiotic stocks based on dH20 should to be filter-sterilized prior to use.
- add less than 5mL of dH20 to the 5mL falcon tube(s).
- multiply the mg of antibiotic listed in the stock column by 5 and add it to the dH20 in the falcon tube.
- dilute the solution to 5mL.
- filter sterilize if not using an antibiotic that is dissolved in ethanol.
- inst. on how to filter sterilize.
- using a P1000, pipette 5 1mL aliquots from the falcon tube into 5 microcentrifuge tubes; store at -20C.
- Store all the aliquots at -20C.
- Tet is light-sensative, so be sure to prevent exposure (i.e. wrap it in tinfoil or keep it in a dark box)