ATCC Medium: 2865 Rhodovulum Iodosum & R. Robiginosum Medium
NaCl…………………………………………….26.40 g
MgCl2 x 6 H20………………………………...5.70 g
MgSO4 x 7 H2O……………………………….6.80 g
CaCl2 x 2 H2O………………………………...1.50 g
KCl……………………………………………...0.66 g
KBr……………………………………………...0.09 g
DI Water………………………………………..965.00 ml
Prepare the medium anaerobically under an atmosphere of N2:CO2 (9:1 vol:vol) and
then add:
NaHCO3 Solution (84 g/l autoclaved under a CO2 atmopshere)………..30.00 ml/l
KH2PO4………………………………………………………………………..0.40 g/l
NH4Cl…………………………………………………………………………..0.25 g/l
Na2S2O3………………………………………………………………………0.50 mM
Trace elements………………………………………………………………..1.00 ml/l
Selenite/tungstate solution…………………………………………………..1.00 ml/l
Vitamin solution……………………………………………………………….1.00 ml/l
Vitamin B12 solution…………………………………………………………1.00 ml/l
Vitamin B1 solution…………………………………………………………..1.00 ml/l
The pH is adjusted to 6.8
Trace Elements
FeSO4 x 7 H2O………………………………….2.10 g
Na2EDTA…………………………………………5.20 g
H3BO3……………………………………………30.00 mg
MnCl2 x 4 H20…………………………………..100.00 mg
CoCl2 x 6 H2O…………………………………..190.00 mg
NiCl2 x 6 H2O……………………………………24.00 mg
CuCl2 x 2 H2O…………………………………..2.00 mg
ZnSO4 x 7 H2O…………………………………144.00 mg
Na2MoO4 x 2 H2O……………………………...36.00 mg
DI Water………………………………………….1000.00 ml
Adjust the pH to 6.0
Selenite/tungsate Solution
Na2SeO3 x 5 H2O………………………………6.00 mg
Na2WO4 x 2 H2O……………………………….8.00 mg
NaOH…………………………………………….200.00 mg
DI Water…………………………………………1000.00 ml
Vitamin Solution
Para-aminobenzoic acid……………………….4.00 mg
Biotin…………………………………………….1.00 mg
Nicotinate……………………………………….10.00 mg
Pantothenate…………………………………..5.00 mg
Pyridoxin……………………………………….15.00 mg
DI Water…………………………………………100.00 ml
Vitamin B12 Solution
Cyanocobalamine………………………………5.00 mg
DI Water…………………………………………100.00 ml
Vitamin B1 Solution
Thiamine…………………………………………10.00 mg
DI Water………………………………………….100.00 ml
To the medium add 5 mM acetate and 10 mM FeSO4 (from a 1 M stock solution kept
under nitrogen). When the iron is added to the medium a white flocculent precipitate
may appear. In the initial stages of growth the precipitate turns orange/brown as the
ferrous ions are oxidised to ferrihydrite. Cultivation may take 2-3 weeks at a distance of
30 cm from a 25 W light bulb.