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1022. CRYPTOANAEROBACTER MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.45 g 0.444 3.213
2 0.40 g 0.394 2.265
3 0.40 g 0.394 7.375
4
(BD Bacto)
5.00 g 4.931 -
5
(BD Bacto)
1.00 g 0.986 -
6 Trace element solution SL-10 2.00 ml - -
7 Selenite-tungstate solution 1.00 ml - -
8
(0.1% w/v)
0.50 ml 4.9e-4 0.002
9 4.00 g 3.945 46.958
10 Supernatant of stationary culture of C. sporogenes 350.00 ml - -
11 Vitamin solution 10.00 ml - -
12 0.08 g 0.079 0.388
13 0.06 g 0.059 0.402
14 650.00 ml 639.876 35518.5
15 , might need strain adjustment 20.00 mg - -
Instructions:
  1. Dissolve ingredients (except bicarbonate, C. sporogenes supernatant, vitamins, magnesium and calcium chloride), adjust pH to 7.0 - 7.5 and boil medium for 1 min, then cool to room temperature under 80% N2 and 20% CO2 gas mixture. Dissolve solid bicarbonate, adjust pH to 7.8, dispense the solution under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add the appropriate amount of sterile and anoxic supernatant of C. sporogenes and complete the medium by adding vitamins (sterilized by filtration), magnesium and calcium chloride from sterile anoxic stock solutions prepared under 100% N2 gas. The final pH of the medium should be 7.5 - 8.0. It may be necessary to add 10 - 20 mg sodium dithionite per liter (e.g. from 5%(w/v) solution, freshly prepared under N2 gas and filter-sterilized), if the medium is not completely reduced after inoculation.
  2. Note: For transfers use 10%(v/v) inoculum.
Equipment needed: Centrifuge Autoclave Anaerobic workbench
Instructions:
  1. Cultivate Clostridium sp. for 5 to 8 days at 37°C in the above medium, but omit 4-hydroxybenzoic acid, replace the C. sporogenes supernatant with distilled water and add after autoclaving 0.30 g/l Na2S x 9 H2O from a sterile anoxic stock solution prepared under 100% N2 gas. Adjust pH of the complete medium to 7.0. Disrupt cells of the grown culture by autoclaving at 121°C for 20 min. Centrifuge autoclaved culture at 18000 x g for 20 min. Discard cell pellet and store the supernatant in screw capped bottles at - 20°C. Before use sterilize the supernatant by autoclaving under 100% N2 gas atmosphere in vials suitable for anaerobic cultivation.
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 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: 7.8
Equipment needed: Anaerobic workbench, 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 652.654 36227.8
Yeast extract 4.931 -
NaHCO3 3.945 46.958
Casamino acids 0.986 -
4-Hydroxybenzoic acid 0.444 3.213
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain

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