This is a cached page for the URL (http://molbiol.ru/eng/protocol/01_05.html). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
   Antibiotics | molbiol.ru   
molbiol.ru - main page.
 > Protocols >  Solutions > Antibiotics.

Antibiotics.

Rambler's Top100



    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 sterilization; or preparation from the sterile water and the sterile powder;
  • store at -20oC in small quantities (about 10 freezing-thawing cycles);
  • instead of H2O it is possible to use 50% glycerol. 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 it very serious: ethanol is a weak catalyst of b-lactam ring disruption. So, solution is unstable.
  • Carbenicillin.

    Conc.1ml5ml10ml
    Carbenicillin50mg/ml50mg250mg0.5g
    H2O 1ml5ml10ml
  • filter sterilization; or preparation 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 sterilization; or preparation 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.

"Practical Molecular Biology" http://molbiol.ru/eng;
e-mail: editor@molbiol.ru

    Supplement.

    People who know! Some information, that you know, but the author did know (or forgot) is missed here. Your colleagues are using this page. Help them, please. Any information, which is helpful for the work are valuable.


    -- do not forget to mark, who is the author of the information! --   

Last modification: 10/11/02



to the beginning

      molbiol.ru - методы, информация и программы для молекулярных биологов    Rambler    Rating@Mail.ru    Be number one