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Main sol. 640a

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Internal Solution-ID: S1365

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.90 g 0.9 16.826
NaCl 0.90 g 0.9 15.4
MgCl2 x 6 H2O 0.40 g 0.4 1.968
KH2PO4 0.75 g 0.75 5.511
K2HPO4 1.50 g 1.5 8.612
Trypticase peptone
(BD BBL) 		2.00 g 2 -
Yeast extract
(OXOID) 		1.00 g 1 -
Trace element solution SL-10 1.00 ml - -
FeCl3 x 6 H2O
(0.1% w/v in 0.2 N HCl) 		2.50 ml 0.003 0.009
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
NaHCO3 2.00 g 2 23.808
Cellobiose 1.00 g 1 2.921
L-Cysteine HCl x H2O 0.50 g 0.5 2.847
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, cellobiose and cysteine. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add bicarbonate and adjust pH to 6.8. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose and cysteine after autoclaving from anoxic stock solutions prepared under 100% N2 gas atmosphere. Cellobiose has to be sterilized by filtration. Adjust pH of the complete medium to 6.5, if necessary.
640a THERMOCLOSTRIDIUM CAENICOLA MEDIUM