MEDIA FOR ANAEROBIC GROWTH 503 



Final Medium 

 Deionized water 800 ml 



Basal salt solution 100 ml 



Biotin (diluted) 3 ml 



DL-malate 2-6 g 



L-glutamate or ammonium sulfate 0.5-2 g 



Phosphate solution (added last) 15 ml 



Deionized water to 1 liter. 



The pH of the medium is 6,8 before autoclaving (15 minutes 

 at 15 Ibs./sq, in.). Immediately after autoclaving, the medium is 

 cloudy but becomes clear upon cooling. 



Incubation temperature: 26-28°C. 



Inoculum Medium 

 (for daily transfers) 

 Same as above, but with 2 g DL-malic acid and 0.5 g am- 

 monium sulfate per liter. The inoculum is grown in completely 

 filled screw-cap tubes. 24-hour-old cells are recommended as 

 inocula. 



B, The Cohen- Bazire, Sistrom and Stanier (1957) medium for Rps. 

 spheroides has also been used for growing R. rubrum. 



Rhodopseudomonas spheroides . 



Stock cultures are maintained as stabs in a medium containing 

 0.3% yeast extract (Difco), 0.2% casamino acids pifco) and 1.5% agar 

 in tap water (see reference given for medium A below). 



A. Semi-synthetic liquid medium. After Cohen- Bazire, Sistrom and 

 Stanier, J. Cellular Co nip. Physiol., 49, 25 (1957). 



Stock Solutions 



1. Potassium phosphate, pH 6.8 



2. Ammonium DL-malate, pH 6.8 



3. Concentrated base: 

 Nitrilotriacetic acid 

 MgS04 



CaCl2 • 2 H2O 

 (NH4)6Mo7024 • 4 H2O 

 FeS04 • 7 H2O 

 Nicotinic acid 

 Thiamine • HCl 

 Biotin 



Metals "44" (see below) 

 Distilled water to 1000 ml. 



