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Main sol. J1203

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Internal Solution-ID: S5322

Equipment needed: Filter Autoclave Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.12 g 0.12 0.815
MgSO4 x 7 H2O 0.19 g 0.19 0.769
KH2PO4 0.45 g 0.449 3.3
K2HPO4 0.45 g 0.449 2.578
NaCl 0.90 g 0.898 15.37
(NH4)2SO4 0.90 g 0.898 6.797
Yeast extract
(BD-Difco) 		2.00 g 1.996 -
Trypticase peptone 2.00 g 1.996 -
n-Butyric acid 0.40 ml - -
iso-Butyric acid 0.40 ml - -
DL-2-Methylbutyric acid 0.20 ml - -
n-Valeric acid 0.20 ml - -
iso-Valeric acid 0.20 ml - -
Resazurin
(0.05%) 		1.00 ml - -
L-Cysteine HCl x H2O 1.00 g 0.998 -
Na2CO3 2.50 g 2.495 23.541
Glucose 2.00 g 1.996 11.079
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except cysteine, carbonate and glucose), adjust pH to 7.0 and sparge medium with 100% CO2 gas for 30-45 min to make it anoxic. Add cysteine and carbonate, then equilibrate the medium with 100% CO2 gas to pH 7.0. Distribute medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add glucose from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
J1203 TREPONEMA SACCHAROPHILUM MEDIUM