Solution A is prepared and autoclaved anaerobically under 80% N2
+ 20% CO2. Solution C is filter-sterilized and gassed for 20
min with N2/CO2. Solutions B, D, E and F are
filter-sterilized and gassed with N2. Solution G is autoclaved
under N2. Solutions B to G are added to the sterile, cooled
solution A in the sequence as indicated. Final pH of the medium should be
7.0. Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5%
(w/v) solution freshly prepared under N2 and filter-sterilized)
may stimulate growth of all strains at the beginning. For transfers use 5
- 10% inoculum. Incubate all strains in the dark.
Solution A
Distilled water
930.0
ml
Resazurin
1.0
mg
CaCl2 x 2 H2O
0.15
g
KCl
0.5
g
MgCl2 x 6 H2O
3.0
g
NaCl
21.0
g
NH4Cl
0.3
g
KH2PO4
0.2
g
Na2SO4
3.0
g
Solution B
Trace elements SL-10 (see medium 116)
1.0
ml
Solution C
Distilled water
50.0
ml
NaHCO3
2.5
g
Solution D
Substrate (see below)
Solution D for E-001657
Distilled water
10.0
ml
Na-acetate x 3 H2O
2.5
g
Solution D for E-001658
Use a gas phase of 80% H2 + 20% CO2 (= substrate).
Cells do not tolerate temperatures higher than 28°C.
Solution E
Vitamin solution (see medium 116)
1.0
ml
Solution F
Na2SeO3 x 5 H2O -solution*
1.0
ml
*3 mg in 1 l of 0.01 M NaOH
Solution G
Distilled water
10.0
ml
Na2S x 9 H2O
0.4
g
Solution A is prepared and autoclaved anaerobically under 80% N2 + 20%
CO2. Solution C is filter-sterilized and gassed for 20 min with N2/CO2.
Solutions B, D, E and F are filter-sterilized and gassed with N2. Solution
G is autoclaved under N2. Solutions B to G are added to the sterile,
cooled solution A in the sequence as indicated. Final pH of the medium
should be 7.0. Addition of 10 - 20 mg sodium dithionite per liter (e.g.
from 5% (w/v) solution freshly prepared under N2 and filter-sterilized)
may stimulate growth of all strains at the beginning. For transfers use 5
- 10% inoculum. Incubate all strains in the dark.
