antibiotics - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki
Ampicillin stock solution (x 1000)
How to prepare Protocol for ampicillin 100 mg/ml stock solution stock (x 1000).
- Weigh ~1 g ampicillin sodium sulfate powder (Fisher 1760-25) in a Falcon 50 mL tube.
- Add water to the 10 mL mark and vortex. Adjust the volume with water if necessary.
- Filter through 0.22 µm membrane with a large syringe.
- Store at -20 in 1 ml aliquots.
Final concentration for E. coli should be 100 µg/ml. This is close to the upper limit recommended by ATCC (see below).
Kanamycin (1000X stock at 50 mg/ml)
Dissolve 1 gram of Kanamycin sulfate in 20 ml distilled water. Vortex to make sure completely dissolved. Sterile by filtering through a 0.2 µm filter. Aliquot and store at -20°C.
Geneticin(G418) stock solution 200 mg/mL (x 1000).
Geneticin (Gibco's brand name) or G418 (generic name) is used to select for yeast that have the KanMX resistance marker. We make G418 stock solution at 200 mg/ml (1000x) and use a final concentration of 200 μg/ml in solid or liquid media. The working solution is stored in 1 mL aliquots marked with a capital "G" at -20°C. Put these tube is the box marked "Antibiotics" in the -20 freezer. Goldstein, A.L., McCusker, J.H., 1999. Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae. Yeast 15, 1541–1553.
Tetracycline 12.5 mg/ml
- Add 125 mg Tetracycline Hydrochloride (MW = 480.898 g/mol) powder to a 15 mL FALCON tube.
- Add 7 mL of Ethanol 99.5 or 96%.
- Add water to 10 mL.
- Aliquot 1 mL per tube and store at -20C.
For XL1-Blue use 12.5 µg/mL Tetracycline.
Antibiotics
Final concentrations generally recommended:
-
100 µg/mL Ampicillin
-
12.5 µg/mL Tetracycline
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50 µg/ml Kanamycin
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200 µg/mL G418
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300 µg/mL HygromycinB
-
200 μg/ml clonNAT
[ atcc.png](atcc_recommendations_antibiotic_concentrations.pdf)