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Solution C

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Internal Solution-ID: S4442

Equipment needed: Autoclave Anaerobic workbench Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4HCO3 0.44 g 6.984 -
NaHCO3 3.73 g 59.206 704.779
Na2SO4 0.20 g 3.175 22.35
Distilled water 50.00 ml - -
1 Prepare all solutions and autoclave under a N2 atmosphere. After cooling, combine the solutions and replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture. Then, aseptically and anaerobically add the following autoclaved or *filter-sterilized solutions to the medium:
Sodium acetate
(1.0 M) 		10.00 ml - -
Vitamin solution
(*) 		1.00 ml - -
Thiamine solution
(*) 		1.00 ml - -
Vitamin B12 solution 0.05 g/L
(*) 		1.00 ml - -
2 Check the final pH of the medium to be around 7.3.
3 Comment: JCM 14587 grows also in a modified medium with benzene and chlorate. For preparation of the modified medium, replace NaNO3 with 1.06 g NaClO3(in 900 ml solution A). After combining all solutions (A, B and C), add 1.0 ml/L of 1.0 M sodium acetate solution (autoclaved), 25 ml/L of 20 mM benzene (water-saturated, autoclaved), and 1 ml/L of each of vitamin solution (see Medium No. 403, filter-sterilized), thiamine solution (see Medium No. 403, filter-sterilized) and vitamin B12 solution (see Medium No. 403, filter-sterilized) to the medium. Otherwise, prepare the modified medium as described above.
J594 ALICYCLIPHILUS DENITRIFICANS MEDIUM