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DSMZ

DSMZ 732 . DEHALOBACTER RESTRICTUS MEDIUM (TCE)

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

Culture medium recipe

Vol.
×

Main sol. 732 #

ml
Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 870 ml - -
Solution B 100 ml - -
Solution C 10 ml - -
Solution D 1 ml - -
Solution E 2 ml - -
Solution F 10 ml - -
Solution G 15 ml - -
1 Sparge solution A with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic serum vials (e.g., 9 ml medium in 50 ml bottle). Pressurize closed bottles with 80% H2 and 20% CO2 gas mixture to 0.5 bar overpressure and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C, D and F are autoclave under 100% N2 gas atmosphere. Solution E is prepared under 100% N2 gas atmosphere and sterilized by filtration. Prepare solution G by filling 13.5 ml hexadecane into a 50 ml serum bottle, then sparge with 100% N2 gas to make it anoxic and autoclave. Add 1.50 ml anoxic autoclaved tetrachloroethene to the sterile anoxic hexadecane solution by syringe. To complete the medium add appropriate amounts of solutions B to F to the sterile solution A in the sequence as indicated. The pH of the medium before inoculation should be at 7.2.
2 Add solution G only after inoculation of the medium!

Solution A #

870
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.65 g 0.747 4.289
NaH2PO4 x H2O 0.17 g 0.195 1.629
Peptone
(BD Bacto) 		0.10 g 0.115 -
Na-acetate 0.46 g 0.529 6.445
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.7e-4 0.002
Distilled water 870.00 ml - -

Solution B #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2CO3 0.27 g 2.7 -
NaHCO3 3.73 g 37.3 444.011
Distilled water 100.00 ml - -

Solution C #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.11 g 11 74.822
MgCl2 x 6 H2O 0.10 g 10 49.188
Distilled water 10.00 ml - -

Solution D #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace element solution 1 ml - -

Solution E #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -
Seven vitamins solution 1 ml - -

Solution F #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.3 g 30 124.905
Distilled water 10.0 ml - -

Solution G #

15
Compound Amount Unit Conc. [g/L] Conc. [mM]
Hexadecane 13.5 ml - -
Tetrachloroethene 1.5 ml - -

Trace element solution #

1000
Solution occurs in 4 other media (first defined in Medium 732).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA 0.5 g 0.5 1.487
FeCl2 x 4 H2O 2.0 g 2 10.06
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
AlCl3 10.0 mg 0.01 0.075
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 1000.0 ml - -
1 First dissolve EDTA in distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add remaining compounds.

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 - -

Seven vitamins solution #

1000
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 13.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle
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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: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 3.7 44.049
K2HPO4 0.645 3.702
Na-acetate 0.456 5.563
Na2S x 9 H2O 0.298 1.239
(NH4)2CO3 0.268 -
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Cultivation metadata from 2 strains BacDive

phyla 2 2 strains
oxygen anaerobe 2 strains
temperature mesophilic 2 strains