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DSMZ

DSMZ 863 . METHANOTHERMOBACTER/METHANOBACTERIUM TRANSDUCTION MEDIUM

This medium was extracted automatically and has not been curated by experts, yet.

Culture medium recipe

Vol.
×

Main sol. 863 #

ml
Equipment needed: Serum bottle Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Basal salts solution 100 ml - -
Trace elements solution 10 ml - -
Sodium carbonate solution 24 ml 59.764 -
Cysteine solutions 5 ml - -
Na2S Solution 5 ml - -
Distilled water
(degassed) 		856 ml - -
1 Anaerobically dispense liquid minimal medium in 10-ml portions into 100-ml serum vials under an atmosphere of N2/CO2 (80:20). Seal vials with butyl rubber stoppers secured with aluminium crimp seals. Sterilize by autoclaving for 15 minutes at 121°C. Prior to inoculation, pressurize bottles to 2 x 102 kPa with H2/CO2. For solidified plating medium, add the following components per liter of liquid minimal medium:
Agar 15 g 14.764 43.897
Titanium (III) citrate solution (Zehnder and Wuhrmann 1976) 16 ml - -
2 Add titanium(III) citrate solution to warm (60°C) agar medium after autoclaving. For preparation of solidified agar medium.

Basal salts solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 68.0 g 68 499.688
NH4Cl 21.2 g 21.2 396.336
Resazurin 10.0 mg 0.01 0.044
Water
(Distilled or deionized) 		1000.0 ml - -

Trace elements solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid
(Aldrich N8407) 		9.550 g 9.55 49.963
MgCl2 x 6 H2O 4.060 g 4.06 19.97
FeCl2 x 4 H2O 0.980 g 0.98 4.929
Na2WO4 x 6 H2O 0.264 g 0.264 -
NiCl2 x 6 H2O 0.118 g 0.118 0.911
Na2Se2O3 0.100 g 0.1 -
Na2MoO4 x 2 H2O 0.024 g 0.024 0.099
CoCl2 x 6 H2O 0.023 g 0.023 0.177
Water
(Distilled or deionized) 		1000.000 ml - -
1 Adjust pH of nitrilotriacetic acid in solution to 7.0 with KOH. Add minerals and adjust the final pH to 7.0.

Na2CO3 solution #

1000
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 106 g 106 1000.11
Water
(degassed) 		1000 ml - -
1 Dissolve 106 g Na2CO3 in 1 liter of degassed H2O. Autoclave in sealed serum bottles under CO2.

Cysteine solutions #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Water 50 ml - -
L-Cysteine HCl 5 g 100 634.437
1 To 50 ml of boiled and cooled distilled or deionized H2O add 5 g of L-cysteine hydrochloride . Adjust pH to 10 with 3N NaOH and store under N2 in a serum bottle sealed with a butyl rubber stopper.

Titanium (III) citrate solution (Zehnder and Wuhrmann 1976) #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Titanium(III) chloride
(Merck 808307, 15% solution) 		5 ml 0.75 -
Sodium citrate
(0.2M) 		50 ml 2.581 10
1 Neutralize pH with a saturated sodium carbonate solution and store in a serum bottle under N2.

Na2S Solution #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Water 50 ml - -
Na2S x 9 H2O 5 g 100 416.351
1 To 50 ml of boiled and cooled distilled or deionized H2O add 5 g of Na2S x 9 H2O. Adjust pH to 10 with 3N NaOH and store under N2 in a serum bottle sealed with a butyl rubber stopper.

Metadata

Last modified: 07.04.22
Source: DSMZ
Medium type: Defined medium
Final pH: 7.0
Equipment needed: Autoclave, Gassing station, Serum bottle
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Gas composition

  • 53 % H2
  • 27 % N2
  • 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: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sodium carbonate solution 59.764 -
Agar 14.764 43.897
KH2PO4 6.693 49.182
NH4Cl 2.087 39.009
L-Cysteine HCl 0.492 3.122
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