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Main sol. 514e

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Internal Solution-ID: S1140

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Peptone
(BD Bacto) 		5.00 g 5 -
Yeast extract
(BD Bacto) 		1.00 g 1 -
NaCl 19.45 g 19.45 332.82
MgCl2
(anhydrous) 		5.90 g 5.9 61.968
Na2SO4 3.24 g 3.24 22.811
KCl 0.55 g 0.55 7.378
NaHCO3 0.16 g 0.16 1.905
KBr 0.08 g 0.08 0.672
SrCl2 34.00 mg 0.034 0.214
H3BO3 22.00 mg 0.022 0.356
Na-silicate 4.00 mg 0.004 0.033
NaF
(0.1% w/v) 		2.40 ml 0.002 0.057
(NH4)NO3
(0.1% w/v) 		1.60 ml 0.002 0.02
Na2HPO4
(0.1% w/v) 		8.00 ml 0.008 0.056
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients except cysteine and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 7.3 - 7.5, if necessary.
514e MARINE BROTH (ANAEROB)