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DSMZ

DSMZ 195d . KNH MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×

Main sol. 195d #

ml
Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 30 ml - -
Solution C 10 ml - -
Solution D 1 ml - -
Solution E 13 ml - -
1 Solution A is sparged with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere in anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is autoclaved under 100% N2 gas. Solutions D and E are prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.1 - 7.4.
2 After inoculation add 1 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.

Solution A #

952
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.151 22.186
KH2PO4 0.20 g 0.21 1.544
NH4Cl 0.30 g 0.315 5.891
NaCl 21.00 g 22.059 377.461
MgCl2 x 6 H2O 3.00 g 3.151 15.5
KCl 0.50 g 0.525 7.045
CaCl2 x 2 H2O 0.15 g 0.158 1.072
Yeast extract 0.50 g 0.525 -
Ascorbic acid 0.10 g 0.105 0.596
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.3e-4 0.002
Distilled water 950.00 ml - -

Solution B #

30
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -

Solution C #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-acetate 0.7 g 70 853.307
Distilled water 10.0 ml - -

Solution D #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -

Solution E #

13
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tris-HCl buffer
(1.0 M solution, pH 8.0) 		7.20 ml - 7.2
Na3-NTA x H2O
(0.5 M solution) 		4.80 ml - 2.4
TiCl3
(15% w/v solution in HCl; Riedel-de Haen) 		0.55 ml 6.346 41.15
1 Sterilize by filtration.

Selenite-tungstate solution #

1000
Solution occurs in 179 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -

Trace element solution SL-10 #

1000
Solution occurs in 266 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 306 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.1 - 7.4
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % H2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 20.875 357.2
Na2SO4 2.982 20.995
MgCl2 x 6 H2O 2.982 14.668
Na2CO3 1.491 14.068
Na-acetate 0.696 8.482
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain