| PubMed I Cell Nature Science Scientific American I HHMI Nobelprize I Weather |
| Dextrose or galactose (40%) | 50 ml |
| Difco yeast nitrogen base (w/o amino acids and ammonium sulfate) | 1.7 g |
| SC-Leu-His-Arg dropout mix | 2 g |
| Sodium Glutamate | 1 g |
| G418 (geneticin) | 300 mg |
| L-Canavanine | 60 mg |
| water | to 1L |
Sterile filter this solution. Geneticin is added at [300 mg/L f.c.] to 65 °C medium. If making plates, just add agar. Arginine has to be omitted from medium for Canavanine to function. Some SGA reporters do not cary Leu+ marker.
Note 1: Ammonium Sulfate in the media, reduce the efficacy of G418. Therefore, when minimal media contain G418, Monosodium Glutamate [1 g/L f.c.] should replace ammonium sulfate as the Nitrogen source (2, 3).
Note 2: The active concentration of Geneticin is written on the bottle and varies from lot to lot (from 500 to 800 μg per mg of total dry weight). The amount used must be adjusted for each lot.
(1) Pan, X et al (2004) Molecular Cell 16, 487-496.
(2) Webster and Dickson (1983). Gene, 26, 243-252.
(3) http://www.openbiosystems.com/