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912. TREPONEMA R8 MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.00 g 0.988 16.908
2 0.50 g 0.494 2.43
3 0.20 g 0.198 1.452
4 0.25 g 0.247 1.418
5 0.30 g 0.296 5.542
6 0.30 g 0.296 3.976
7 15.00 mg 0.015 0.101
8 Trace element solution SL-10 1.00 ml - -
9 Selenite-tungstate solution 1.00 ml - -
10
(SIGMA)
2.10 g 2.075 -
11 0.90 g 0.889 4.936
12 0.19 g 0.188 -
13
(BD Bacto)
0.19 g 0.188 -
14
(0.1% w/v)
0.50 ml 4.9e-4 0.002
15 2.50 g 2.47 29.407
16 Clarified rumen fluid 50.00 ml - -
17 Vitamin solution 10.00 ml - -
18 0.40 g 0.395 2.25
19 0.40 g 0.395 1.646
20 950.00 ml 937.052 52014.2
Instructions:
  1. Dissolve ingredients (except bicarbonate, rumen fluid, vitamins, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate, adjust pH to 7.2, distribute medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add rumen fluid, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins should be sterilized by filtration. Check pH of complete medium and adjust to 7.2, if necessary.
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 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 366 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.0 mg 0.002 0.008
2 2.0 mg 0.002 0.005
3 10.0 mg 0.01 0.049
4 5.0 mg 0.005 0.015
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 0.01
8 0.1 mg 1.0e-4 6.3e-5
9 5.0 mg 0.005 0.036
10 5.0 mg 0.005 0.024
11 1000.0 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:  Bacteria
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

  • 100 % N2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 948.879 52670.7
NaHCO3 2.47 29.407
MOPS 2.075 -
NaCl 0.988 16.908
D-Glucose 0.889 4.936
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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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