抗生素溶液配制

互联网2013-09-05

7213

Working concentrations and stock solutions
Preparation of stock solutions

Ampicillin
Carbenicillin
GE antibiotic mix
Tetracycline
Chloramphenicol

Supplement.

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Working concentrations and stock solutions.

Antibiotics	Stock solution	Working concentration:
Strict control	Weak control
mg/ml:		µg/ml	dilution	µg/ml	dilution
Ampicillin	50 (H₂ undefined)	-20^o C	20	0.4 µl/ml	60-125	1.2-2.5 µl/ml
Carbenicillin	50 (H₂ O, 50%EtOH)	-20^o C	20	0.4 µl/ml	60-200	1.2-4 µl/ml
Chloramphenicol	34 (EtOH, MetOH)	-20^o C	25	0.7 µl/ml	170	5 µl/ml
D-cycloserinundefined	10 (0.1M NaP pH8.0)	+4^o C	200	20 µl/ml
Gentamycin	10 (H₂ O)	-20^o C	15	1.5 µl/ml
Hygromycin B	10 (H₂ O)	-20^o C	10	1 µl/ml	400	40 µl/ml
Kanamicin	50 (H₂ O)	+4^o C- -20^o C	10	0.2 µl/ml	50	1 µl/ml
Kasugamycin	10 (H₂ O)	-20^o C	1000	100 µl/ml
Nalidixic acid	5 (H₂ O, pH to 11 /NaOH)	-20^o C	15	3.0 µl/ml
Neomycin	10 (H₂ O)	-20^o C	100	10 µl/ml	800	80 µl/ml
Rifampiciundefined~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M34_~AMetOH~B~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M~F20~Ksup~Mo~K~Hsup~M_C~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M150~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M4.4_µl/ml
Spectinomycin	10 (H₂ O)	-20^o C	100	10 µl/ml
Streptomicin	50 (H₂ O)	-20^o C	10	0.2 µl/ml	50	1 µl/ml
Tetracyclinundefined*~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M5_~F_12.5~Kbr_~H~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~AEtOH~E_70~7EtOH~B~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M~F20~Ksup~Mo~K~Hsup~M_C~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M10~K~Hfont~M~K~Htd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~Ktd~M~2~1~0~0~0~0~0~0~0~0~0~0~0~0~0~Kfont~M2_µl/ml	50 100	10 µl/ml 20 µl/ml

1ml

5ml

10ml

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

store at -20^o C in small aliquots (avoid more than 10 freezing-thawing cycles);

it is possible to use 50% glycerol instead of H₂ O. The advantage - solution is liquid at -20^o C.

some laboratories use 96% (or 50%) EtOH for ampicillin solubilization. There are two advantages in this method: (i) solution is liquid at -20^o C, (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.	1ml	5ml	10ml
Carbenicillin	50mg/ml	50mg	250mg	0.5g
H₂ O		1ml	5ml	10ml

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

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

200 µg/ml for pETpfu.

GE antibiotic mix (50x).

	Conc.	40ml
Cefotaxime 25µg/ml	25µg/ml	1mg
Ceftriaxone 25µg/ml	25µg/ml	1mg
Potassium phosphate (pH 6.0) 0.1M		40ml

filter sterilize; or prepare from the sterile KPO₄ and the sterile powder;

store at -20⁰ C, in small quantities (about 10 freezing-thawing cycles).

Tetracycline.

	Conc.	1ml	5ml	10ml
Tetracycline	5mg/ml	5mg	25mg	50mg
EtOH		1ml	5ml	10ml

solution in EtOH, store at -20^o C;

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

Chloramphenicol.

	Conc.	1ml	5ml	10ml
Chloramphenicol	34mg/ml	34mg	170mg	340mg

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

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