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Main sol. 701

Internal Solution-ID: S1452

Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 1 18.695
MgCl2 x 6 H2O 7.0 g 7 34.431
MgSO4 x 7 H2O 9.6 g 9.6 38.949
CaCl2 x 2 H2O 0.5 g 0.5 3.401
KCl 3.8 g 3.8 50.973
K2HPO4 x 3 H2O 0.4 g 0.4 2.297
NaCl 10.0 g 10 171.116
NaHCO3 3.0 g 3 35.711
Na2CO3 1.0 g 1 9.435
Na2S x 9 H2O 0.5 g 0.5 2.082
Yeast extract 1.0 g 1 -
L-Cysteine 0.5 g 0.5 4.127
Resazurin 1.0 mg 1.0e-3 0.004
Substrate
(glucose, chitin etc.)
5.0 g 5 -
Modified Wolin's mineral solution II 1.0 ml - -
Distilled water 1000.0 ml - -
1 pH 7.2
2 Prepare the medium without the Na2S, Na2CO3, NaHCO3, L-cysteine and substrate. Boil the medium and cool under a stream of nitrogen. Add the Na2CO3 and NaHCO3 . Adjust the pH to 7.2. If chitin is used as the substrate it must be added before autoclaving. Autoclave the medium and add the Na2S (from a sterile, neutralized stock solution prepared under nitrogen), L-cysteine (from a stock solution prepared under nitrogen), and the glucose or N-acetylglucosamine (if used as substrate) from sterile stock solution prepared under nitrogen. Check the final pH.
701 HALOANAEROBACTER CHITINOVORANS MEDIUM