520
520 .
RUMINOCLOSTRIDIUM CELLULOLYTICUM (CM3) MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Medium for Caldanaerobacter subterraneus subsp. yonseiensis DSM 13777
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
75 °C
Main sol. 520 #
ml
Equipment needed: Filter
Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| (NH4)2SO4 | 1.30 | g | 1.296 | 9.809 | |
| KH2PO4 | 1.50 | g | 1.496 | 10.99 | |
| K2HPO4 x 3 H2O | 2.90 | g | 2.891 | 16.6 | |
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
1.25 | ml | 0.001 | 0.004 | |
| Trace element solution SL-10 | 1.00 | ml | - | - | |
| Yeast extract | 2.00 | g | 1.994 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
| MgCl2 x 6 H2O | 0.20 | g | 0.199 | 0.981 | |
| CaCl2 x 2 H2O | 75.00 | mg | 0.075 | 0.509 | |
| Cellobiose | 6.00 | g | 5.982 | 17.476 | |
|
Cellulose
, MN 301
optional |
10.00 | g | 9.97 | - | |
| Na2CO3 | 1.50 | g | 1.496 | 14.11 | |
| L-Cysteine HCl x H2O | 0.50 | g | 0.499 | 2.838 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except magnesium chloride, calcium chloride, cellobiose, cysteine and carbonate, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add magnesium chloride, calcium chloride and cellobiose from anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2. Cellobiose has to be sterilized by filtration. Prior to inoculation add cysteine from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH to 7.2. | |||||
| 2 Note: Some strains can be adapted to cellulose as substrate using 10.00 g/l cellulose powder MN 301 (MACHEREY-NAGEL). | |||||
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. | |||||
Metadata
| Last modified: | 04.04.22 |
| Source: | DSMZ |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | 7.2 |
| Equipment needed: | Autoclave, Filter, Gassing station, Hungate tubes |
| Edit in Medium builder | |
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: | 15 |
| Strains with modifications: | 4 |
Caldanaerobacter subterraneus subsp. yonseiensis DSM 13777
No modifications necessary for this strain.
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
|
Cellulose optional |
9.97 | - |
| Cellobiose | 5.982 | 17.476 |
| K2HPO4 x 3 H2O | 2.891 | 16.6 |
| Yeast extract | 1.994 | - |
| Na2CO3 | 1.496 | 14.11 |
| Show more | ||
