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734. METHANOBREVIBACTER CURVATUS MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 Clarified rumen fluid 400.00 ml - -
2 1.00 g 0.996 17.043
3 0.50 g 0.498 6.68
4 0.40 g 0.398 1.96
5 0.10 g 0.1 0.677
6 0.30 g 0.299 5.586
7 0.20 g 0.199 1.464
8 0.15 g 0.149 1.052
9
(BD Bacto)
0.50 g 0.498 -
10
(OXOID)
0.50 g 0.498 -
11
(BD Difco)
2.00 g 1.992 -
12 Trace element solution SL-10 1.00 ml - -
13 Selenite-tungstate solution 1.00 ml - -
14
(0.1% w/v)
0.50 ml 5.0e-4 0.002
15 2.00 g 1.992 18.795
16
(SIGMA)
2.10 g 2.092 -
17 Seven vitamins solution 1.00 ml - -
18 0.16 g 0.159 1.033
19 600.00 ml 596.538 33112.9
Instructions:
  1. Dissolve ingredients (except carbonate, MOPS buffer, vitamins and DTT), bring medium to the boil, then cool to room temperature under 80% H2 and 20% CO2 gas mixture. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add MOPS buffer adjusted to pH 7.2, vitamins and DTT from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.2, if necessary.
Solution occurs in 32 other media (first defined in medium 1310 )
Equipment needed: Centrifuge Filter Autoclave
Instructions:
  1. Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 minand then centrifuged at 27,000 g for 20 min The supernatant is made anoxic by sparging with 100% N2 gas for 15 min dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at - 20°C.
Solution occurs in 161 other media (first defined in medium 385 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.5 g 0.5 12.501
2 3.0 mg 0.003 0.011
3 4.0 mg 0.004 0.012
4 1000.0 ml 998.207 55408.8
Solution occurs in 86 other media (first defined in medium 503 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 100 mg 0.1 0.063
2 80 mg 0.08 0.583
3 20 mg 0.02 0.082
4 200 mg 0.2 1.625
5 100 mg 0.1 0.21
6 300 mg 0.3 1.459
7 200 mg 0.2 0.536
8 1000 ml 998.207 55408.8
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, Centrifuge, Filter, Hungate tubes

Associated strains

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

Gas composition

  • 20 % CO2
  • 80 % N2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 599.511 33277.9
MOPS 2.092 -
Na2CO3 1.992 18.795
Nutrient broth 1.992 -
NaCl 0.996 17.043
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain

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