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Main sol. J556

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Internal Solution-ID: S4370

Equipment needed: Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 0.100 g 0.1 1.708
MgCl2 x 6 H2O 0.040 g 0.04 0.196
KCl 0.050 g 0.05 0.669
CaCl2 x 2 H2O 0.015 g 0.015 0.102
Na2SO4 0.016 g 0.016 0.112
Distilled water 993.000 ml - -
1 Mix components and autoclave. After cooling, add the following solutions (autoclaved or filter-sterilized) to the medium:
HEPES buffer
(pH 7.0, 2 M) 		5.000 ml - -
1 M Phosphate buffer
(pH 7.2, 1 M) 		0.150 ml - -
KNO3
(0.09 M) 		0.544 ml - -
FeCl2 solution 1.000 ml - -
Trace element solution 1.000 ml - -
Vitamin solution
(filter-sterilized) 		1.000 ml - -
2 Distribute the culture medium into culture tubes (leave >80% volume as a gas phase) and seal with butyl rubber stoppers.
3 Finally, replace the gas phase with 20% methane and 5% CO2 in air (by volume).
J556 METHYLOSOMA DIFFICILE MEDIUM