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Main sol. 806

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Internal Solution-ID: S1637

Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.33 4.427
MgCl2 x 2 H2O 2.70 g 2.7 20.573
MgSO4 x 7 H2O 3.40 g 3.4 13.794
NH4Cl 0.25 g 0.25 4.674
CaCl2 x 2 H2O 0.14 g 0.14 0.952
K2HPO4 0.14 g 0.14 0.804
Na2SeO3 1.00 µg 1.0e-6 5.8e-6
NiCl2 x 6 H2O 1.00 µg 1.0e-6 7.7e-6
NaHCO3 1.00 g 1 11.904
NaCl 18.00 g 18 308.008
Resazurin 1.00 mg 1.0e-3 0.004
L-Cysteine HCl x H2O 0.50 g 0.5 2.847
Na2S x 9 H2O 0.50 g 0.5 2.082
Yeast extract 1.00 g 1 -
Trypticase 1.00 g 1 -
Sulphur 5.00 g 5 -
Modified Wolin's mineral solution II 10.00 ml - -
Wolin's vitamin solution 10.00 ml - -
Distilled water 980.00 ml - -
1 Prepare the medium without NaHCO3, cysteine hydrochloride, Na2S x 9 H2O, and vitamin solution. Boil the medium and cool under N2. Add the NaHCO3 to the cooled medium and adjust the pH to about pH 7.2. Sterilise the medium at 100°C for 3 hours on 3 consecutive days. Reduce the medium by adding 10% neutralised sodium sulphide and cysteine hydrochloride from sterile stock solutions. The vitamins are added from a sterile (filter sterilised) stock solution prepared under nitrogen. The final pH is 7.2.
806 THERMOCOCCUS MEDIUM