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JCM 747 . FRESHWATER MEDIUM WITH TOLUENE

Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.

Culture medium recipe

Vol.
×

Solution A #

36
Equipment needed: Anaerobic workbench Filter Autoclave
1 First dissolve iron(III) citrate in 800 ml distilled water and adjust pH to 7.0 with 10 N NaOH. Then, add the remaining minerals, bring volume to 970 ml and autoclave under a N2 atmosphere. Add aseptically and anaerobically the following filter-sterilized solutions:
Compound Amount Unit Conc. [g/L] Conc. [mM]
Iron(III) citrate 12.00 g 333.333 1360.88
NaCl 1.00 g 27.778 475.321
KCl 0.50 g 13.889 186.303
MgCl2 x 6 H2O 0.40 g 11.111 54.653
NH4Cl 0.25 g 6.944 129.827
KH2PO4 0.20 g 5.556 40.824
CaCl2 x 2 H2O 0.10 g 2.778 18.895
Na2SO4 1.42 mg 0.039 0.278
Sodium acetate x 3 H2O 1.36 g 37.778 -
NaHCO3
(8.5%) 		30.00 ml - -
FeCl2 solution 1.00 ml - -
Trace element solution (see Medium No. 187 1.00 ml - -
Vitamin solution 1.00 ml - -
Thiamine solution 1.00 ml - -
Vitamin B12 solution 0.05 g/L 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
2 Distribute the medium into sterile culture vessels (e.g., 50 ml per 120 ml bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with sterile butyl rubber stoppers. Then add per 50 ml the following solutions from anaerobic stocks:
FeCl2
(100 mM) 		0.50 ml - -
Toluene 5.00 �l - -
3 Check the final pH of the medium to be around 7.0.

FeCl2 solution #

1000
Solution occurs in 87 other media (first defined in Medium J187).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(7.7 M, 25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
Distilled water 990.0 ml - -

Vitamin solution #

100
Solution occurs in 14 other media (first defined in Medium J403).
Compound Amount Unit Conc. [g/L] Conc. [mM]
p-Aminobenzoic acid 4 mg 0.04 0.292
Biotin 1 mg 0.01 0.041
Nicotinic acid 10 mg 0.1 0.812
DL-Calcium pantothenate 5 mg 0.05 0.105
Pyridoxine hydrochloride 10 mg 0.1 -
Sodium phosphate buffer
(10 mM, pH 7.1) 		100 ml - -

Thiamine solution #

100
Solution occurs in 10 other media (first defined in Medium J403).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Thiamine HCl 10 mg 0.1 -
Sodium phosphate buffer
(25 mM, pH 3.4) 		100 ml - -

Vitamin B12 solution 0.05 g/L #

100
Solution occurs in 18 other media (first defined in Medium J403).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 5 mg 0.05 0.037
Distilled water 100 ml - -

Selenite-tungstate solution #

1000
Solution occurs in 71 other media (first defined in Medium J431).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.4 g 0.4 10.001
Na2SeO3 x 5 H2O 6.0 mg 0.006 0.023
Na2WO4 x 2 H2O 8.0 mg 0.008 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 22.02.22
Source: JCM
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 7.0
Equipment needed: Anaerobic workbench, Autoclave, Filter
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Associated strains

All strains for this medium: 2

Cultivation metadata from 2 strains BacDive

pH 6 - 8 2 strains
phyla 2 2 strains
oxygen anaerobe 2 strains
temperature mesophilic 2 strains