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Main sol. J198

Internal Solution-ID: S3863

Equipment needed: Anaerobic workbench Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.75 g 0.736 5.408
K2HPO4 0.75 g 0.736 4.226
MnCl2 x 4 H2O 0.36 g 0.353 -
NH4Cl 1.00 g 0.981 18.347
NaHCO3 4.00 g 3.925 46.727
Trace mineral solution 9.00 ml - -
Trace vitamins 10.00 ml - -
Na2S x 9 H2O 0.50 g 0.491 2.043
L-Cysteine HCl x H2O 0.50 g 0.491 -
Resazurin 1.00 mg 9.8e-4 0.004
Distilled water 1000.00 ml - -
1 Mix ingredients, except NaHCO3, Trace vitamins, Na2S x 9H2O and L-cysteine x HCl x H2O, and autoclave under a H2-CO2 (80:20) atmosphere. Filter-sterilize NaHCO3 as 8% solution and Trace vitamins while gassing with N2. Separately autoclave Na2S x 9H2O and L-cysteine x HCl x H2O as 5% solutions under a N2 atmosphere. Aseptically and anaerobically add the NaHCO3 solution and Trace vitamins to the medium under a H2-CO2 (80:20) atmosphere. Prior to inoculation, add sterile the Na2S x 9H2O and L-cysteine x HCl solutions, and adjust pH to 7.4. Pressurize inoculated bottles to 200 kPa H2-CO2 (80:20).
J198 JCM MEDIUM No. 198