Materials and Reagents

1. Freshly harvested Streptomyces spores in water or mycelium in 50 mM MOPS buffer (pH 7)
   
2. Tryptic Soy Broth (TSB) (Sigma-Aldrich, catalog number: 22092 )
   
3. 3-(N-morpholino)propanesulfonic acid (MOPS) buffer (Roth North America)
   
4. Sulfanilic acid (Sigma-Aldrich, catalog number: 251917 )
   
5. N-(1-naphthyl)-ethylenediamine dihydrochloride (Roth North America)
   
6. Hydrochloric acid (Roth North America)
   
7. Sodium hydroxide pellets (Roth North America)
   
8. Soya flour (from local supermarket)
   
9. D-mannitol (Sigma-Aldrich catalogue number 63560 )
   
10. Agar-agar (Kobe I) (Roth North America)
    
11. Sulfanilic acid solution (see Recipes)
    
12. N-(1-naphthyl)-ethylenediamine solution (see Recipes)
    
13. MOPS-buffer (see Recipes)
    
14. SFM agar (see Recipes)
    

Equipment

1. Cotton wool (standard issue from local pharmacy)
   
2. 0.22 µm pore-size filters (PVDF) (Roth North America)
   
3. 30 °C rotary shaker
   
4. Baffled Erlenmeyer flasks (500 ml) (Glasgerätebau Ochs, Laborfachhandel e. K., catalog number: 100500 )
   
5. Standard-sized plastic Petri dishes for bacterial growth and spore preparation
   
6. Gas-tight glass Hungate tubes (16 ml) for anaerobic work with butyl rubber septa (Glasgerätebau Ochs, Laborfachhandel e. K., catalog number: 1020471 )
   
7. Cooled table-top centrifuge (e.g. Eppendorf)
   
8. Cylinder of pure nitrogen gas (e.g. Linde or local supplier)
   
9. Needles 0.6 mm Gauge (B. Braun Melsungen AG)
   
10. Water bath or heating block (Biometra)
    
11. Spectrophotometer measuring absorption in the visible range
    

Procedure