抗生素溶液配制-试验方案

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Working concentrations and stock solutions
Preparation of stock solutions

Ampicillin
Carbenicillin
GE antibiotic mix
Tetracycline
Chloramphenicol

Supplement.

About the recalculation of recipes for the arbitrary volume:

Working concentrations and stock solutions.

AntibioticsStock solutionWorking concentration:Strict controlWeak controlmg/ml:         µg/mldilutionµg/mldilutionAmpicillin50 (H₂O*)-20^oC200.4 µl/ml60-1251.2-2.5 µl/mlCarbenicillin50
(H₂O, 50%EtOH)-20^oC200.4 µl/ml60-2001.2-4 µl/mlChloramphenicol34
(EtOH, MetOH)-20^oC250.7 µl/ml1705 µl/mlD-cycloserine*10
(0.1M NaP pH8.0)+4^oC20020 µl/ml  Gentamycin10 (H₂O)-20^oC151.5 µl/ml  Hygromycin B10 (H₂O)-20^oC101 µl/ml40040 µl/mlKanamicin50 (H₂O)+4^oC-
-20^oC100.2 µl/ml501 µl/mlKasugamycin10 (H₂O)-20^oC1000100 µl/ml  Nalidixic acid5
(H₂O, pH to 11 /NaOH)-20^oC153.0 µl/ml  Neomycin10 (H₂O)-20^oC10010 µl/ml80080 µl/mlRifampicin**34 (MetOH)-20^oC1504.4 µl/ml  Spectinomycin10 (H₂O)-20^oC10010 µl/ml  Streptomicin50 (H₂O)-20^oC100.2 µl/ml501 µl/mlTetracycline***5 - 12.5
(EtOH, 70%EtOH)-20^oC102 µl/ml50
10010 µl/ml
20 µl/ml

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

Preparation of stock solutions.

Ampicillin.

Conc.1ml5ml10mlAmpicillin50mg/ml50mg250mg0.5gH₂O 1ml5ml10ml

filter sterilize; or prepare from the sterile water and the sterile powder;

store at -20^oC 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 -20^oC.

some laboratories use 96% (or 50%) EtOH for ampicillin solubilization. There are two advantages in this method: (i) solution is liquid at -20^oC, (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.1ml5ml10mlCarbenicillin50mg/ml50mg250mg0.5gH₂O 1ml5ml10ml

filter sterilize; or prepare from the sterile water and the sterile powder;

store at -20^oC in small quantities (about 10 freezing-thawing cycles);

200 µg/ml for pETpfu.

GE antibiotic mix (50x).

Conc.40mlCefotaxime 25µg/ml25µg/ml1mgCeftriaxone 25µg/ml25µg/ml1mgPotassium 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.1ml5ml10mlTetracycline5mg/ml5mg25mg50mgEtOH 1ml5ml10ml

solution in EtOH, store at -20^oC;

Mg⁺⁺- antagonist, for the selection it is necessary to use Mg-free medium.

Chloramphenicol.

Conc.1ml5ml10mlChloramphenicol34mg/ml34mg170mg340mg

store dry powder at 4^oC, solution at -20^oC (in EtOH);

BL21(DE3)pLyzS - 34µg/ml -> 1µl for 1ml of medium.

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抗生素溶液配制
点击： 作者： 来源： 日期：2006-11-07 本站论坛