Culture medium recipe
Vol.
×
Main sol. 385 #
ml
Equipment needed: Gassing station
Hungate tubes
Autoclave
Filter
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Solution A | 932 | ml | - | - | |
Solution B | 1 | ml | - | - | |
Solution C | 20 | ml | - | - | |
Solution D | 10 | ml | - | - | |
Solution E | 10 | ml | - | - | |
Solution F | 10 | ml | - | - | |
Solution G | 10 | ml | - | - | |
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min ), then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solutions B and G are autoclaved separately under 100% N2 gas. | |||||
2 Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solutions D, E and F are prepared under 100% N2 gas and sterilized by filtration. The pyrocatechol stock solution has to be prepared always freshly prior to use. To complete the medium appropriate amounts of the solutions B to G are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be at 6.9 - 7.1. | |||||
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the beginning. For transfers use 10 - 20% inoculum. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2SO4 | 3.00 | g | 3.219 | 22.662 |
KH2PO4 | 0.20 | g | 0.215 | 1.577 |
NH4Cl | 0.30 | g | 0.322 | 6.018 |
NaCl | 7.00 | g | 7.511 | 128.52 |
MgCl2 x 6 H2O | 1.30 | g | 1.395 | 6.861 |
KCl | 0.50 | g | 0.536 | 7.196 |
CaCl2 x 2 H2O | 0.15 | g | 0.161 | 1.095 |
Selenite-tungstate solution | 1.00 | ml | - | - |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - |
Distilled water | 930.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Trace element solution SL-10 | 1 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2CO3 | 1 | g | 50 | 471.752 |
Distilled water | 20 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na-benzoate | 0.4 | g | 40 | 277.579 |
Distilled water | 10.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Pyrocatechol | 0.06 | g | 6 | 54.491 |
HCl
(1 N) |
0.06 | ml | - | - |
Distilled water | 10.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Wolin's vitamin solution | 10 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2S x 9 H2O | 0.4 | g | 40 | 166.54 |
Distilled water | 10.0 | ml | - | - |
Solution occurs in 173
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 | - | - |
Solution occurs in 262
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. |
Solution occurs in 351
other media (first defined in Medium 141).
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Biotin | 2.0 | mg | 0.002 | 0.008 |
Folic acid | 2.0 | mg | 0.002 | 0.005 |
Pyridoxine hydrochloride | 10.0 | mg | 0.01 | 0.049 |
Thiamine HCl | 5.0 | mg | 0.005 | 0.015 |
Riboflavin | 5.0 | mg | 0.005 | 0.013 |
Nicotinic acid | 5.0 | mg | 0.005 | 0.041 |
Calcium D-(+)-pantothenate | 5.0 | mg | 0.005 | - |
Vitamin B12 | 0.1 | mg | 1.0e-4 | 7.4e-5 |
p-Aminobenzoic acid | 5.0 | mg | 0.005 | 0.036 |
(DL)-alpha-Lipoic acid | 5.0 | mg | 0.005 | 0.024 |
Distilled water | 1000.0 | ml | - | - |
No comments for this medium.