Antibiotics - ANTIBIOTICS A. Antibiotic Concentrations....

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ANTIBIOTICS A. Antibiotic Concentrations. Antibiotic Abbreviation Final concentration 1 Stock solution 2 Rich media Minimal media Na Ampicillin 3 Amp or Ap 30 μg/ml 15 μg/ml 9 mg/ml Chloramphenicol Cam or Cm 20 μg/ml 5 μg/ml 6 mg/ml Gentamycin SO 4 Gen or Gm 10μg/ml ---- 3 mg/ml Kanamycin SO 4 4 Kan or Km 50 μg/ml 125 μg/ml 15 mg/ml Tetracycline HCl 5 Tet or Tc 20 μg/ml 10 μg/ml 6 mg/ml Timentin --- 80 μg/ml --- 24 mg/ml Trimethoprim Tm 200 μg/ml --- 15 mg/ml Spectinomycin Spc or Sp 100 μg/ml 50 μg/ml 50 mg/ml Streptomycin SO 4 6 Str or Sm --- 2 mg/ml --- 1 The solid form of the antibiotics can be added directly to sterilized media that has been cooled to approximately 55°C. If kept at 4°C tetracycline, chloramphenicol, and streptomycin plates are usually good for several months, but kanamycin plates and ampicillin plates may only last for several weeks. A sensitive and resistant control should always be included each time antibiotic plates are used. 2 For liquid media or for a few plates, a stock solution of the antibiotics can be prepared and stored at -20°C. All the antibiotic stock solutions can be prepared in sterile dH 2 O except chloramphenicol which can be dissolved in dimethylformamide and timentin which can be dissolved in 70% ethanol. The stock solution is 300x the concentration of antibiotic required in rich medium. An average petrie dish contains about 30 ml medium, so spread 0.1 ml on rich plates. 3 High level expression of ß-lactamase can destroy ampicillin in the medium surrounding the Amp R colonies, allowing Amp s satellite colonies to grow. Satellite colonies can be decreased by using 2x ampicillin when selecting for plasmids. Alternatively, Timentin prevents growth of
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This note was uploaded on 06/08/2011 for the course MICROB 101 taught by Professor Bio during the Spring '11 term at Rutgers.

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Antibiotics - ANTIBIOTICS A. Antibiotic Concentrations....

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