154. DESULFOVIBRIO SULFODISMUTANS MEDIUM

Solution 1:

KH2PO4 0.2 g

NH4Cl 0.3 g

NaCl 1.0 g

MgCl2 6 H2O 0.4 g

KCl 0.5 g

CaCl2 2 H2O 0.15 g

Distilled water 920.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO3 2.5 g

Distilled water 50.0 ml

Solution 4:

Na-acetate 3 H2O 0.3 g

Distilled water 10.0 ml

Solution 5:

D(+)-Biotin 10.0 mg

Ca-D(+)-Pantothenate 50.0 mg

Distilled water 1.0 ml

Solution 6:

Na2S 9 H2O 0.4 g

Distilled water 10.0 ml

Solution 7:

Na2S2O3 (0.5 M) 10.0 ml

pH 7.5-8.0 (adjust with NaOH)

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl2 4 H2O 1.5 g

ZnCl2 70.0 mg

MnCl2 4 H2O 100.0 mg

H3BO3 6.0 mg

CoCl2 6 H2O 190.0 mg

CuCl2 2 H2O 2.0 mg

NiCl2 6 H2O 24.0 mg

Na2MoO4 2 H2O 36.0 mg

Distilled water 990.0 ml

pH 7.1-7.4

Solution 1 is prepared and autoclaved anaerobically at under 80% N2 + 20% CO2 at 121C for 15 min. Solutions 2, 4, 6 and 7 are gassed with N2 and autoclaved separately at 121C for 15 min. Solution 3 (gassed with N2 + CO2) and solution 5 (gassed with N2) are filter-sterilized. Solutions with 2 to 7 are added to the sterile, cooled solution 1 in the sequence as indicated. When growth has started feed culture again with same amount of solution 7. After a further two days repeat feeding once more.

 

Updated 21.06.2021