Medium data
Search for medium no. = [862].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
862 STEROIDOBACTER MEDIUM
| NaCl | 1.0 | g |
| MgCl2·6H2O | 0.4 | g |
| KH2PO4 | 0.2 | g |
| KCl | 0.5 | g |
| NH4Cl | 0.25 | g |
| CaCl2·2H2O | 0.15 | g |
| Na2SO4 | 0.07 | g |
| NaNO3 | 0.42 | g |
| Distilled water | 1.0 | L |
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.
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.
After autoclaving, add (per 10 ml) 0.3 ml NaHCO3 solution (84 g/L, autoclaved under a CO2 atmosphere), 1 µl Trace element solution SL-10 (see Medium No. 433), 1 µl Selenite-tungstate solution (see below) and 1 µl 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:
| NaOH | 0.5 | g |
| Na2SeO3·5H2O | 3.0 | mg |
| Na2WO4·2H2O | 4.0 | mg |
| Distilled water | 1.0 | L |
Vitamin solution:
| Vitamin B12 | 50.0 | mg |
| Pantothenic acid | 50.0 | mg |
| Riboflavin | 50.0 | mg |
| Pyridoxamine·HCl | 10.0 | mg |
| Biotin | 20.0 | mg |
| Folic acid | 20.0 | mg |
| Nicotinic acid | 25.0 | mg |
| Nicotinamide | 25.0 | mg |
| α-Lipoic acid | 50.0 | mg |
| p-Aminobenzoic acid | 50.0 | mg |
| Thiamine·HCl·2H2O | 50.0 | mg |
| Distilled water | 1.0 | L |
Copyright © 2025 Microbe Division (JCM) - All Rights Reserved