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Main sol. J187

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Internal Solution-ID: S3845

Equipment needed: Autoclave Anaerobic workbench Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
CaCl2 x 2 H2O 0.44 g 0.439 2.987
MgCl2 x 6 H2O 0.70 g 0.699 3.436
NaCl 0.50 g 0.499 8.539
NaHCO3 0.80 g 0.798 9.504
FeCl2 solution 1.00 ml - -
Trace element solution 1.00 ml - -
Trace vitamins 10.00 ml - -
Yeast extract
(BD-Difco) 		0.20 g 0.2 -
Bacto peptone
(BD-Difco) 		5.00 g 4.99 -
Sulfur
(powder) 		10.00 g 9.98 311.195
Na2S x 9 H2O 0.50 g 0.499 2.078
Resazurin 1.00 mg 1.0e-3 0.004
Distilled water 990.00 ml - -
1 Mix ingredients, except yeast extract, peptone, vitamins, sulfur and Na2S x 9H2O, and autoclave under a N2-CO2 (80:20) atmosphere. Separately autoclave yeast extract and peptone as 10% solutions and Na2S x 9H2O as a 5% solution under a N2 atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Filter-sterilize NaHCO3 as an 8% solution and vitamin solution while gassing with N2. Aseptically and anaerobically add yeast extract, peptone, NaHCO3 and vitamin solutions, and sulfur to the medium under a N2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution, and adjust pH to 6.2-6.4 with sterile 1 N H2SO4.
J187 DESULFUROCOCCUS AMYLOLYTICUS MEDIUM