Medium data

Search for medium no. = [747].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

747   FRESHWATER MEDIUM WITH TOLUENE

Solution A:
Iron(III) citrate  12.0     g
NaCl  1.0     g
KCl  0.5     g
MgCl2·6H2O  0.4     g
NH4Cl  0.25    g
KH2PO4  0.2     g
CaCl2·2H2O  0.1     g
Na2SO4  1.42    mg
Sodium acetate·3H2O  1.36    g
First dissolve iron(III) citrate in 800 ml distilled water and adjust pH to 7.0 with 10 N NaOH. Then, add the remaining minerals, bring volume to 970 ml and autoclave under a N2 atmosphere. Add aseptically and anaerobically the following filter-sterilized solutions:
8.5% NaHCO3 solution  30.0     ml
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187  1.0     ml
Vitamin solution (see Medium No. 403)  1.0     ml
Thiamine solution (see Medium No. 403)  1.0     ml
Vitamin B12 solution (see Medium No. 403)  1.0     ml
Selenite-tungstate solution (see Medium No. 431)  1.0     ml
Distribute the medium into sterile culture vessels (e.g., 50 ml per 120 ml bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with sterile butyl rubber stoppers. Then add per 50 ml the following solutions from anaerobic stocks:
100 mM FeCl2 solution  0.5     ml
Toluene  5.0     µl
Check the final pH of the medium to be around 7.0.