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Antibiotic solutions




    About the recalculation of recipes for the arbitrary volume:

    Working concentrations and stock solutions.

    AntibioticsStock solution Working concentration:
    Strict controlWeak control
    mg/ml:         µg/ml dilutionµg/mldilution
    Ampicillin50 (H2O*)-20oC200.4 µl/ml60-1251.2-2.5 µl/ml
    Carbenicillin50
    (H2O, 50%EtOH)
    -20oC200.4 µl/ml60-2001.2-4 µl/ml
    Chloramphenicol34
    (EtOH, MetOH)
    -20oC250.7 µl/ml1705 µl/ml
    D-cycloserine*10
    (0.1M NaP pH8.0)
    +4oC20020 µl/ml  
    Gentamycin10 (H2O)-20oC151.5 µl/ml  
    Hygromycin B10 (H2O)-20oC101 µl/ml40040 µl/ml
    Kanamicin50 (H2O)+4oC-
    -20oC
    100.2 µl/ml501 µl/ml
    Kasugamycin10 (H2O)-20oC1000100 µl/ml  
    Nalidixic acid5
    (H2O, pH to 11 /NaOH)
    -20oC153.0 µl/ml  
    Neomycin10 (H2O)-20oC10010 µl/ml80080 µl/ml
    Rifampicin**34 (MetOH)-20oC1504.4 µl/ml  
    Spectinomycin10 (H2O)-20oC10010 µl/ml  
    Streptomicin50 (H2O)-20oC100.2 µl/ml501 µl/ml
    Tetracycline***5 - 12.5
    (EtOH, 70%EtOH)
    -20oC102 µl/ml50
    100
    10 µl/ml
    20 µl/ml

    * - Unstable solution, prepare just before use;
    ** - Light sensitive;
    *** - Light sensitive, Mg2+ - inhibitor;



    Preparation of stock solutions

    Ampicillin

    Conc.1ml5ml10ml
    Ampicillin50mg/ml50mg250mg0.5g
    H2O 1ml5ml10ml
  • filter sterilize; or prepare from the sterile water and the sterile powder;
  • store at -20oC in small aliquots (avoid more than 10 freezing-thawing cycles);
  • it is possible to use 50% glycerol instead of H2O. The advantage - solution is liquid at -20oC.
  • some laboratories use 96% (or 50%) EtOH for ampicillin solubilization. There are two advantages in this method: (i) solution is liquid at -20oC, (ii) the solution is self sterilizing. But the disadvantage is very serious: ethanol is a weak catalyst of ß-lactam ring disintegration. So, solution is unstable.
  • Carbenicillin

    Conc.1ml5ml10ml
    Carbenicillin50mg/ml50mg250mg0.5g
    H2O 1ml5ml10ml
  • filter sterilize; or prepare from the sterile water and the sterile powder;
  • store at -20oC in small quantities (about 10 freezing-thawing cycles);
  • 200 µg/ml for pETpfu.
  • GE antibiotic mix (50x)

    Conc.40ml
    Cefotaxime 25µg/ml25µg/ml1mg
    Ceftriaxone 25µg/ml25µg/ml1mg
    Potassium phosphate (pH 6.0) 0.1M 40ml
  • filter sterilize; or prepare from the sterile KPO4 and the sterile powder;
  • store at -200C, in small quantities (about 10 freezing-thawing cycles).
  • Tetracycline

    Conc.1ml5ml10ml
    Tetracycline5mg/ml5mg25mg50mg
    EtOH 1ml5ml10ml
  • solution in EtOH, store at -20oC;
  • Mg++- antagonist, for the selection it is necessary to use Mg-free medium.
  • Chloramphenicol

    Conc.1ml5ml10ml
    Chloramphenicol34mg/ml34mg170mg340mg
  • store dry powder at 4oC, solution at -20oC (in EtOH);
  • BL21(DE3)pLyzS - 34µg/ml -> 1µl for 1ml of medium.


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Last modification: 30/07/13

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