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DSMZ

DSMZ 1022 . CRYPTOANAEROBACTER MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 1022 #

ml
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
4-Hydroxybenzoic acid 0.45 g 0.448 3.245
K2HPO4 0.40 g 0.398 2.287
NH4Cl 0.40 g 0.398 7.448
Yeast extract
(BD Bacto) 		5.00 g 4.98 -
Casamino acids
(BD Bacto) 		1.00 g 0.996 -
Trace element solution SL-10 2.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 4.00 g 3.984 47.425
C. sporogenes supernatant 350.00 ml - -
MgCl2 x 6 H2O 0.08 g 0.08 0.392
CaCl2 x 2 H2O 0.06 g 0.06 0.406
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 650.00 ml - -
1 Dissolve ingredients (except bicarbonate, C. sporogenes supernatant, magnesium chloride, calcium chloride, and vitamins), 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 magnesium chloride, calcium chloride and vitamins (sterilized by filtration) from sterile anoxic stock solutions prepared under 100% N2 gas. The final pH of the medium should be 7.5 - 8.0.
2 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.
3 Note: For transfers use 10% (v/v) inoculum.

Trace element solution SL-10 #

1000
Solution occurs in 265 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Selenite-tungstate solution #

1000
Solution occurs in 178 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -

C. sporogenes supernatant #

1000
1 Cultivate Clostridium sp. DSM 754 for 5 to 8 days at 37°C in the medium 1022, 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.

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: 10.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.5 - 8.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % 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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Yeast extract 4.98 -
NaHCO3 3.984 47.425
Casamino acids 0.996 -
4-Hydroxybenzoic acid 0.448 3.245
NH4Cl 0.398 7.448
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Cultivation metadata from 1 strains BacDive

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