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Making Antibiotics and Stocks
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Antibiotics

Updated: 11.11.21 KL

 

We use certain antibiotics in the lab for selection of bacteria. We have several standard ones. They need to be made at a certain concentration in a certain solvent

Below are the details. Final concentration you would use depends on application.

Store all at -20C.

This list is adapted from :https://barricklab.org/twiki/bin/view/Lab/ProtocolsAntibioticStockSolutions

Reagent Abbreviation Colour Line

Stock Conc.

(mg/mL)

Working Conc.

(ug/mL)

Dilution Solvent Notes

Kanamycin

Kan / K Red -- 50 50 1000X Water General use
Ampicillin Amp / A Blue -- 100 100 1000X Water General use, unstable, slowly switching to Carb
Carbenecillin Carb / C Blue -- 100 100 1000X Water Replacing Amp usage
Chloramphenicol Chl Black -- 25 25 1000X EtOH  
Tetracycline Tet / T Green -- 12.5 12.5 1000X EtOH  
Gentamycin Gent / G Black

--

--

10 10 1000X Water Not used a lot, typically for Bacmid selection
Spectinomycin Spec     50 50   Water  

 


 

Stocks

Below is a summary of our common stocks and their concentrations and pH.

They should always be stocks and should be filtered 0.22 um

Chemical Notes
5 M NaCl  
1 M HEPES, pH 7.5 HEPES does not dissolve readily, it needs to be adjusted with NaOH to dissolve. Takes time
2.5 M Imidazole, pH 7.5 Imidazole is basic when dissolved, we adjust it to make sure that people do not accidetally basify their proteins
0.5 M EDTA, pH 8.0 EDTA needs to be pHed like HEPES
1 M TRIS, pH 7.5 or 8 We purchase this from the Lavarie
10X PBS We purchase this from the Lavarie, final pH can be 6.8 or 7.4
0.5 M MES, pH 5 or 6 MES is not highly soluble and is light sensitive, store away from light
1 M DTT store -20
1 M IPTG store -20
0.5 M TCEP must be neutralized or else extremely acidic. not stable in phosphate buffers.

 

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