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Main sol. 323

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Internal Solution-ID: S599

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.12 g 0.12 0.816
MgSO4 x 7 H2O 0.19 g 0.19 0.771
KH2PO4 0.45 g 0.45 3.307
K2HPO4 0.45 g 0.45 2.584
NaCl 0.90 g 0.9 15.4
(NH4)2SO4 0.90 g 0.9 6.811
Yeast extract
(OXOID) 		2.00 g 2 -
Trypticase peptone
(BD BBL) 		2.00 g 2 -
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 - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
L-Cysteine HCl x H2O 1.00 g 1 5.694
Na2CO3 2.50 g 2.5 23.588
D-Glucose 2.00 g 2 11.101
Agar , for solid medium
(BD Bacto) 		12.00 g 12 35.679
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 the cysteine and carbonate, then equilibrate the medium with the CO2 gas to pH 7.0. Distribute medium under same 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.
2 For solid medium add 12.00 g/l agar (BD Bacto).
323 TREPONEMA SACCHAROPHILUM MEDIUM