Stock, conc | Preparation & Storage | Volumes, Tubes | Location | |
---|---|---|---|---|
10× T4 ligase buffer | Thawed, precipitates resuspended with vortexing and/or brief 37° incubation. | 50 µL aliquots | Rack 1.5 | |
10 mM dNTPs | Thawed | 20 µL aliquots | Rack 1.5 | |
Antibiotics | ||||
chl25 (mg/mL) | Ethanol, -20°; stable for ≥1 year. | 15 mL | Freezer 1 door | |
amp100 | 40% ethanol, -20°; seems to be stable for ≥6 mo for agar | 15 mL | Freezer 1 door | |
kan50 | Water, 0°; stable for ≥1 year. | 15 mL | Freezer 0 door | |
spt50 | Water, 0°; stable for ≥1 year. | 15 mL | Freezer 0 door | |
gen | Water, -20°; stable for… | 1 mL | Rack 1.5 |
Antibiotics
Antibiotic | Target | Stock Concentration | Solvents | Notes | Rich Medium | ||||
---|---|---|---|---|---|---|---|---|---|
Abbr* | Relative | m/V | 1× c, Multi-Copy; Range | 1× c, Single-Copy | |||||
Chloramphenicol | ribosome | chl | 1000× | 25 mg/mL (formerly 34 mg/mL) | 100% Ethanol | 25 or 34 mg/L (3) | ≥25–340 mg/L (5) | 10 mg/L (1) or 5 mg/L (6) | |
Ampicillin·Na | peptidoglycan | amp | 1000× | 100 mg/mL | 40% ethanol or water | 40% EtOH keeps it liquid at -20°C. | 100 mg/L (3) | ≥100–200 mg/L | 50 mg/L (1) |
Kanamycin B·SO4 | ribosome | kan | 1000× | 50 mg/mL | 35% DMSO or Water | Insoluble in 40% ethanol. Precipitates from 35% DMSO. | 50 mg/L (3) | ≥50–500 mg/L (5) | 20 mg/L (1) |
Spectinomycin·2HCl·5H2O | ribosome | spt | 1000× | 50 mg/mL | Water | Precipitates from 35% DMSO. | 50 mg/L (3) | ≥50–500 mg/L (5) | 20 mg/L (1) |
Streptomycin·SO 4 | ribosome | str | 1000× | 100 mg/mL | Water | ||||
Trimethoprim | FolA (DHFR) | tmp | 1000× | 15 mg/mL | DMSO or 1% acetic acid | Insoluble in water or ethanol. Exogenous thymine can lead to evolution of resistance. | 15 mg/L? | 10 mg/L (2) | |
Apramycin·SO 4 | ribosome | apr | 1000× | ? | Water | ? | ? | ||
Gentamicin·SO 4 | ribosome | gen | 1000× | 10 mg/mL? | Water | 10 mg/L (8) | 10 mg/L (1) | ||
Erythromycin | ribosome | ery | |||||||
Tetracycline·HCl | ribosome | tet | 1000× | 10 mg/mL | Water or ? | Light-sensitive. | 10 mg/L (3) | 15 mg/L (1) | |
Zeocin (Phleomycin D1 · Cu2+) | DNA | zeo | 1000× | 25 mg/mL | Water | Use with LB-Lennox (not >5 g/L NaCl), pH 7.5. Light-sensitive. | 25 mg/L (7) | 25–50 mg/L (7) | |
Hygromycin B | ribosome | hyg | 1000× | 200 mg/mL | Water | 200 mg/L (3) |
References
* Follow American Society of Microbiology AAC abbreviation guidelines when possible
- Poteete et al., Biotechniques (2006)
- Kittleson et al., J Biol Eng (2011)
- Addgene
- Qiagen
- Sara Molinari, unpublished data, p15A with cat, aphA1, aadA.
- Addgene: Marionette Sensor Collection
- Zeocin | Thermo Fisher Scientific