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JCM 827 . DEHALOCOCCOIDES MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. J827 #

ml
Equipment needed: Filter Autoclave Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.000 g 0.946 16.189
MgCl2 x 6 H2O 0.410 g 0.388 1.908
NH4Cl 0.270 g 0.255 4.775
KCl 0.520 g 0.492 6.599
CaCl2 x 2 H2O 0.015 g 0.014 0.097
KH2PO4 0.200 g 0.189 1.39
FeCl2 solution 1.000 ml - -
Trace element solution 1.000 ml - -
Selenite-tungstate solution 1.000 ml - -
Sodium acetate 0.410 g 0.388 4.729
Resazurin 0.250 mg 2.4e-4 0.001
Distilled water 1000.000 ml - -
1 Mix components thoroughly, bring to a boil and cool down under a N2 atmosphere. Add the following solutions from anaerobic stocks, then dispense the medium into culture vessels leaving 40 to 50% volume of a gas phase (e.g., 30 ml of the medium in a 50 ml serum bottle) and autoclave under a N2 atmosphere:
L-Cysteine HCl x H2O
(5%) 		1.000 ml - -
Na2S x 9 H2O
(5%) 		1.000 ml - -
2 After cooling, replace the gas phase with a N2-CO2-H2 (8:1:1, v/v) gas mixture. Add per liter the following filter-sterilized solutions from anaerobic stocks:
NaHCO3
(8%) 		10.000 ml - -
MOPS
(pH 7.4, 1.0 M) 		20.000 ml - -
25 mM Ti(III) NTA solution
(25 mM) 		20.000 ml - -
Vitamin solution 1.000 ml - -
3 To complete the medium, add per liter the following filter-sterilized chlorosolvent:
Trichloroethylene
(10 mg/ml in methanol) 		0.700 ml - -
4 Check final pH to be about 7.2.

Vitamin solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 10 mg 0.01 0.041
Folic acid 2 mg 0.002 0.005
Pyridoxine hydrochloride 250 mg 0.25 -
Thiamine HCl 20 mg 0.02 -
Riboflavin 5 mg 0.005 0.013
Nicotinic acid 100 mg 0.1 0.812
DL-Calcium pantothenate 10 mg 0.01 0.021
Vitamin B12 25 mg 0.025 0.018
p-Aminobenzoic acid 50 mg 0.05 0.365
Lipoic acid 5 mg 0.005 0.024
Distilled water 1000 ml - -

25 mM Ti(III) NTA solution #

300
Equipment needed: Anaerobic workbench Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 9.6 g 32 167.417
Water
(anaerobic ) 		300.0 ml - -
1 To prepare anaerobic water, boil 1.0 L distilled water and cool down under a stream of oxygen-free N2 gas. Mix components under N2 gas. Adjust pH to 9.0 with NaOH. Slowly add 9.6 ml of 20% TiCl3 to the chelator solution. Keep the pH above 2.0 by the addition of anaerobic saturated Na2CO3 solution. Adjust pH to 7.0 with Na2CO3 solution, adjust pH to 7.0 and bring the final volume to 500 ml with the anaerobic water. Filter-sterilize and keep the solution under a N2 gas atmosphere.

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 - -

Trace element solution #

1000
Solution occurs in 89 other media (first defined in Medium J187).
Compound Amount Unit Conc. [g/L] Conc. [mM]
ZnCl2 70 mg 0.07 0.514
MnCl2 x 4 H2O 100 mg 0.1 0.505
H3BO3 6 mg 0.006 0.097
CoCl2 x 6 H2O 190 mg 0.19 1.463
CuCl2 x 2 H2O 2 mg 0.002 0.012
NiCl2 x 6 H2O 24 mg 0.024 0.185
Na2MoO4 x 2 H2O 36 mg 0.036 0.149
Distilled water 1000 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
Medium type: Defined medium
Final pH: 9.0
Equipment needed: Anaerobic workbench, Autoclave, Filter
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Associated strains

All strains for this medium: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 0.946 16.189
Nitrilotriacetic acid 0.605 3.168
KCl 0.492 6.599
Sodium acetate 0.388 4.729
MgCl2 x 6 H2O 0.388 1.908
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