DSMZ 441 . DIAZOTROPHIC MEDIUM (RBA)

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Culture medium recipe

Vol.
×
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 953 ml - -
Solution B 50 ml - -
Standard vitamin solution 5 ml - -
1 Sterilize solution A separately at 121°C for 15 min., cool to 50°C and then mix aseptically with filter-sterilized solution B and 5.0 ml of filter-sterilized standard vitamin solution (see medium 428).
2 RBA is an ammonium-free medium which has successfully been used for the isolation, growth and purity check of a broad spectrum of nitrogen fixing bacteria (Ref. 3363). For microaerophilic nitrogen-fixing bacteria use semisolid medium with 0.3% end concentration of agar and incubate the liquid cultures under 10% (v/v) air and 90% (v/v) N2.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.10 g 0.105 0.771
K2HPO4 0.90 g 0.944 5.422
NaCl 0.10 g 0.105 1.796
CaCl2 x 2 H2O 0.10 g 0.105 0.714
MgSO4 x 7 H2O 0.10 g 0.105 0.426
Na2MoO4 x 2 H2O 5.00 mg 0.005 0.022
NaVO3 x H2O 5.00 mg 0.005 0.037
MnSO4 x H2O 5.00 mg 0.005 0.031
FeSO4 x 7 H2O 0.01 g 0.01 0.038
Yeast extract 0.05 g 0.052 -
Trace element solution SL-6 3.00 ml - -
Distilled water 950.00 ml - -
Agar , if required 15.00 g 15.74 46.799
1 Adjust pH to 7.3.
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Disodium succinate 1 g 20 123.419
DL-Malate 2 g 40 298.307
Na-pyruvate 1 g 20 181.745
D-Mannitol 2 g 40 219.575
D-Glucose 2 g 40 222.025
Distilled water 50 ml - -
1 Adjust pH to 7.3.
Solution occurs in 55 other media (first defined in Medium 27).
Compound Amount Unit Conc. [g/L] Conc. [mM]
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
MnCl2 x 4 H2O 0.03 g 0.03 0.152
H3BO3 0.30 g 0.3 4.851
CoCl2 x 6 H2O 0.20 g 0.2 1.54
CuCl2 x 2 H2O 0.01 g 0.01 0.059
NiCl2 x 6 H2O 0.02 g 0.02 0.154
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
Distilled water 1000.00 ml - -
Solution occurs in 3 other media (first defined in Medium 428).
Equipment needed: Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Riboflavin 10.0 mg 0.1 0.266
Thiamine-HCl x 2 H2O 50.0 mg 0.5 1.339
Nicotinic acid 50.0 mg 0.5 4.061
Pyridoxine hydrochloride 50.0 mg 0.5 2.431
Calcium pantothenate 50.0 mg 0.5 1.049
Biotin 0.1 mg 1.0e-3 0.004
Folic acid 0.2 mg 0.002 0.005
Vitamin B12 1.0 mg 0.01 0.007
Distilled water 100.0 ml - -
1 Filter sterilize (0.2 µm pore-size) and store at 4°C.

Metadata

Last modified: 20.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Autoclave, Filter
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Associated strains

All strains for this medium: 49
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Agar 14.881 44.245
DL-Malate 1.984 14.797
D-Glucose 1.984 11.013
D-Mannitol 1.984 10.892
Na-pyruvate 0.992 9.015
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Cultivation metadata from 49 strains BacDive

phyla 2 49 strains
oxygen obligate aerobe 1 strain
temperature mesophilic 49 strains