Editing Antibiotics

Working concentrations and stock solutions. 

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| colspan="2" rowspan="2" | Stock solution 
| align="CENTER" colspan="4" | Working concentration:
|- bgcolor="eeeeee"
| colspan="2" | Strict control 
| colspan="2" | Weak control
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| mg/ml: 
| 
| µg/ml 
| dilution 
| µg/ml 
| dilution
|- bgcolor="ffffff"
| Ampicillin 
| 50 (H<sub>2</sub>O*) 
| -20<sup>o</sup>C 
| 20 
| 0.4 µl/ml 
| 60-125 
| 1.2-2.5 µl/ml
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| Carbenicillin 
| 50<br>(H<sub>2</sub>O, 50%EtOH) 
| -20<sup>o</sup>C 
| 20 
| 0.4 µl/ml 
| 60-200 
| 1.2-4 µl/ml
|- bgcolor="ffffff"
| Chloramphenicol 
| 34<br>(EtOH, MetOH) 
| -20<sup>o</sup>C 
| 25 
| 0.7 µl/ml 
| 170 
| 5 µl/ml
|- bgcolor="ffffff"
| D-cycloserine* 
| 10<br>(0.1M NaP pH8.0) 
| 4<sup>o</sup>C 
| 200 
| 20 µl/ml 
| 
| 
|- bgcolor="ffffff"
| Gentamycin 
| 10 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 15 
| 1.5 µl/ml 
| 
| 
|- bgcolor="ffffff"
| Hygromycin B 
| 10 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 10 
| 1 µl/ml 
| 400 
| 40 µl/ml
|- bgcolor="ffffff"
| Kanamicin 
| 50 (H<sub>2</sub>O) 
| 4<sup>o</sup>C-<br>-20<sup>o</sup>C 
| 10 
| 0.2 µl/ml 
| 50 
| 1 µl/ml
|- bgcolor="ffffff"
| Kasugamycin 
| 10 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 1000 
| 100 µl/ml 
| 
| 
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| Nalidixic acid 
| 5<br>(H<sub>2</sub>O,<font size="-1">pH to 11 /NaOH</font>) 
| -20<sup>o</sup>C 
| 15 
| 3.0 µl/ml 
| 
| 
|- bgcolor="ffffff"
| Neomycin 
| 10 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 100 
| 10 µl/ml 
| 800 
| 80 µl/ml
|- bgcolor="ffffff"
| Rifampicin** 
| 34 (MetOH) 
| -20<sup>o</sup>C 
| 150 
| 4.4 µl/ml 
| 
| 
|- bgcolor="ffffff"
| Spectinomycin 
| 10 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 100 
| 10 µl/ml 
| 
| 
|- bgcolor="ffffff"
| Streptomicin 
| 50 (H<sub>2</sub>O) 
| -20<sup>o</sup>C 
| 10 
| 0.2 µl/ml 
| 50 
| 1 µl/ml
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| Tetracycline*** 
| 5 - 12.5<br>(EtOH, 70%EtOH) 
| -20<sup>o</sup>C 
| 10 
| 2 µl/ml 
| 50<br>100 
| 10 µl/ml<br>20 µl/ml
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Unstable solution, prepare just before use;<br> ** - Light sensitive;<br> *** - Light sensitive, Mg<sup>2 </sup> - inhibitor; 

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Preparation of stock solutions. Ampicillin. 

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| Conc. 
| 1ml 
| 5ml 
| 10ml 
| 
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| Ampicillin 
| 50mg/ml 
| 50mg 
| 250mg 
| 0.5g 
| 
|- bgcolor="ffffff"
| H<sub>2</sub>O 
| 
| 1ml 
| 5ml 
| 10ml 
| 
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*filter sterilization; or preparation from the sterile water and the sterile powder; 
*store at -20<sup>o</sup>C in small quantities (about 10 freezing-thawing cycles); 
*instead of H<sub>2</sub>O it is possible to use 50% glycerol. The advantage - solution is liquid at -20<sup>o</sup>C. 
*some laboratories use 96% (or 50%) EtOH for ampicillin solubilization. There are two advantages in this method: (i) solution is liquid at -20<sup>o</sup>C, (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. 

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| Conc. 
| 1ml 
| 5ml 
| 10ml 
| 
|- bgcolor="ffffff"
| Carbenicillin 
| 50mg/ml 
| 50mg 
| 250mg 
| 0.5g 
| 
|- bgcolor="ffffff"
| H<sub>2</sub>O 
| 
| 1ml 
| 5ml 
| 10ml 
| 
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*filter sterilization; or preparation from the sterile water and the sterile powder; 
*store at -20<sup>o</sup>C in small quantities (about 10 freezing-thawing cycles); 
*200 µg/ml for pETpfu.

Chloramphenicol. 

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| Conc. 
| 1ml 
| 5ml 
| 10ml 
| 
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| Chloramphenicol 
| 34mg/ml 
| 34mg 
| 170mg 
| 340mg 
| 
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*store dry powder at 4<sup>o</sup>C, solution at -20<sup>o</sup>C (in EtOH); 
*BL21(DE3)pLyzS - 34µg/ml -&gt; 1µl for 1ml of medium.

GE antibiotic mix (50x). 

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| Conc. 
| 40ml 
| 
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| Cefotaxime 25µg/ml 
| 25µg/ml 
| 1mg 
| 
|- bgcolor="ffffff"
| Ceftriaxone 25µg/ml 
| 25µg/ml 
| 1mg 
| 
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| KPO<sub>4</sub>(pH 6.0) 0.1M 
| 
| 40ml 
| 
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*filter sterilization; or preparation from the sterile KPO<sub>4</sub> and the sterile powder; 
*store at -20<sup>0</sup>C, in small quantities (about 10 freezing-thawing cycles).

Tetracycline. 

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| Conc. 
| 1ml 
| 5ml 
| 10ml 
| 
|- bgcolor="ffffff"
| Tetracycline 
| 5mg/ml 
| 5mg 
| 25mg 
| 50mg 
| 
|- bgcolor="ffffff"
| EtOH 
| 
| 1ml 
| 5ml 
| 10ml 
| 
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*solution in EtOH, store at -20<sup>o</sup>C. 
*Mg<sup></sup>- antagonist, for the selection it is necessary to use Mg-free medium.

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[[Category:Buffer_Recipe]]