Common antibiotic stock solutions for laboratory use. When preparing antibiotics stocks, I generally prepare in 15 mL falcon tubes, then aliquot into 1.5 mL tubes for long-term storage.
Use 100 µL of stock per 100 mL medium for appropriate concentration. For additional details for usage see here.
Reducing pH improves dissolving of Ampicillin in ddH2O
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 100 µg/mL for selection of common ampr (β-lactamase – ble) vectors.
Carbenicillin (100 mg/mL) in ddH2O stock
Carbenicillin
1
g
Sterile ddH2O to
10
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 100 µg/mL for selection of common ampr (β-lactamase – ble) vectors.
Carbenicillin can be subsituted for Ampicillin as ble confers resistance to both. Carb is a semi-synthetic form of Amp and is generally more stable.
Chloramphenicol (25 mg/ml) in EtOH stock
Chloramphenicol
250
mg
Ethanol (100%)
10
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 25 µg/mL for selection.
Gentamycin (100 mg/mL) in ddH2O stock
Gentamycin
1
g
Sterile ddH2O to
10
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 25 µg/mL for selection.
Kanamycin (50 mg/mL) in ddH2O stock
Kanamycin sulfate
0.50
g
Sterile ddH2O to
10
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 50 µg/mL for selection.
Paromomycin (20 mg/mL) in ddH2O stock
Paromomycin
1
g
Sterile ddH2O to
50
mL
Filter sterilise (0.22 µm filter).
Aliquot 1000 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 375 µL per 100 mL media for a final working concentration of 15 µg/mL for selection of paror (aminoglycoside 3′-phosphotransferase type VIII – AphVIII) vectors
Rifampicin (200 mg/mL) in EtOH stock
Rifampicin
200
mg
Ethanol (100%)
1
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 200 µg/mL for Agrobacterium selection.
Spectinomycin (100 mg/mL) in ddH2O
Spectinomycin
1
g
Sterile ddH2O to
10
mL
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 100 µg/mL for selection.
Tetracycline (10 mg/mL) in 50:50 EtOH:ddH2O stock
Tetracycline
1
g
Ethanol (100%)
50
mL
Sterile ddH2O to
100
mL
Dissolve in 50% Ethanol.
Filter sterilise (0.22 µm filter).
Aliquot 500 µl into autoclaved 1.5 mL tubes. Store at -20 °C, protected from light.
Use 100 µL per 100 mL media for final concentration of 10 µg/mL for selection.
