Name: STEROIDOBACTER MEDIUM
Creator: MediaDive
License: http://creativecommons.org/licenses/by/4.0/

Main sol. J862 #

ml

Equipment needed: Autoclave Anaerobic workbench Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	1.00	g	1	17.112
MgCl₂ x 6 H₂O	0.40	g	0.4	1.968
KH₂PO₄	0.20	g	0.2	1.47
KCl	0.50	g	0.5	6.707
NH₄Cl	0.25	g	0.25	4.674
CaCl₂ x 2 H₂O	0.15	g	0.15	1.02
Na₂SO₄	0.07	g	0.07	0.493
NaNO₃	0.42	g	0.42	4.941
Distilled water	1000.00	ml	-	-
1 For growth with testosterone, dissolve testosterone in acetone (20 mg/ml) and dispense portions of the solution, e.g., 0.1 ml portions for 10 ml medium, to anaerobic culture tubes. Let the solvent evaporate to dryness. Dispense 10 ml portions of medium in the culture tubes and stream with N₂-CO₂ (80:20, v/v). Close the vessels and autoclave. Treat the vessels in an ultrasonic bath to detach and suspend the testosterone.
2 For growth with heptanoate, omit testosterone. Sterilize the gassed medium in anaerobic vessels by autoclaving. Add 2.5 mM heptanoate from a 20-fold filter-sterilized anaerobic stock solution after cooling.
3 After autoclaving, add (per 10 ml) 0.3 ml NaHCO₃ solution (84 g/L, autoclaved under a CO₂ atmosphere), 1 ï¿½l Trace element solution SL-10 (see Medium No. 433), 1 ï¿½l Selenite-tungstate solution (see below) and 1 ï¿½l Vitamin solution (see below). Adjust pH to 7.2. Incubate the cultures in the dark for up to 4 weeks. Briefly shake the cultures once per day.

Selenite-tungstate solution #

1000

Solution occurs in 178 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	-	-

1000

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Vitamin B₁₂	50	mg	0.05	0.037
Pantothenic acid	50	mg	0.05	0.228
Riboflavin	50	mg	0.05	0.133
Pyridoxamine hydrochloride	10	mg	0.01	-
Biotin	20	mg	0.02	0.082
Folic acid	20	mg	0.02	0.045
Nicotinic acid	25	mg	0.025	0.203
Nicotinamide	25	mg	0.025	0.205
alpha-lipoic acid	50	mg	0.05	0.242
p-Aminobenzoic acid	50	mg	0.05	0.365
Thiamine-HCl x 2 H₂O	50	mg	0.05	-
Distilled water	1000	ml	-	-

Last modified:	22.02.22
Source:	JCM
Medium type:	Defined medium
Final pH:	7.2
Equipment needed:	Anaerobic workbench, Autoclave, Filter

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