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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 |
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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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