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DSMZ

DSMZ 1280 . METHANOREGULA (PEAT) MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 1280 #

ml
Equipment needed: Autoclave Serum bottle Gassing station Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 954 ml - -
Solution B 10 ml - -
Solution C 13 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
1 Dissolve ingredients of solution A, adjust pH to 5.0 with 0.1 N HCl and sparge with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense solution A under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 50 ml bottles) and autoclave. Solutions B - F are prepared under 100% N2 gas atmosphere and sterilized by filtration. Prior to inoculation complete the medium by adding appropriate amounts of solutions B to F to the sterile solution A in the sequence as indicated.
2 Adjust pH of the complete medium to 5.1 with a sterile anoxic solution of 1 N HCl. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3 Note: Use 10-20% (v/v) as inoculum.

Solution A #

954
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 27.0 mg 0.028 0.529
KH2PO4 14.0 mg 0.015 0.108
KCl
(0.1% w/v) 		2.0 ml 0.002 0.028
Na-acetate 17.0 mg 0.018 0.217
Trace elements solution 1.0 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.2e-4 0.002
Distilled water 950.0 ml - -

Solution B #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Homo-PIPES
(SIGMA) 		1.58 g 158 -
NaOH
(pellets) 		0.18 g 18 450.034
Distilled water 10.00 ml - -
1 Adjust pH of buffer to 5.5.

Solution C #

13
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH
(0.38 M) 		7.20 ml 0.109 2.736
Nitrilotriacetate
(0.5 M, neutralized with 10 N NaOH) 		4.80 ml 0.459 2.4
TiCl3
(15% w/v in HCl, RIEDEL-de HAEN) 		0.55 ml 6.346 41.15

Solution D #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.2 g 20 -
Distilled water 10.0 ml - -

Solution E #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-Mercaptoethanesulfonate
(coenzyme M) 		0.07 g 7 49.226
Distilled water 10.00 ml - -

Solution F #

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

Trace elements solution #

1010
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.00 ml 2.475 67.889
FeCl2 x 4 H2O 1.34 g 1.327 6.673
CoCl2 x 6 H2O 24.00 mg 0.024 0.183
ZnCl2 75.00 mg 0.074 0.545
H3BO3 19.00 mg 0.019 0.304
NiCl2 x 6 H2O 24.00 mg 0.024 0.183
Na2MoO4 x 2 H2O 24.00 mg 0.024 0.098
MnSO4 x 4 H2O 26.00 mg 0.026 0.115
MgSO4 x 7 H2O 1.56 g 1.545 6.267
CaCl2 x 2 H2O 2.34 g 2.317 15.759
CuSO4 x 5 H2O 9.00 mg 0.009 0.036
AlK(SO4)2 x 12 H2O 3.45 g 3.416 7.201
Distilled water 1000.00 ml - -
1 First dissolve ferrous chloride in the HCl, then dilute in water and dissolve the other salts. Finally make up to 1000 ml.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 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: 03.05.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 5.1
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Homo-PIPES 1.569 -
Yeast extract 0.199 -
TiCl3 0.082 0.531
2-Mercaptoethanesulfonate 0.07 0.489
NH4Cl 0.027 0.501
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Cultivation metadata from 1 strains BacDive

pH 4 - 6 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain