265. ECTOTHIORHODOSPIRA VARIABILIS MEDIUM

NaCl 50.00 g

Na2CO3 5.00 g

NaHCO3 15.00 g

KCl 0.10 g

K2HPO4 0.50 g

MgCl2 × 6 H2O 0.20 g

NH4Cl 0.50 g

Na2S × 9 H2O 0.50 g

Na2S2O3 × 5 H2O 0.50 g

Yeast extract 0.10 g

Na acetate 0.50 g

Trace element solution SL-10 (see below) 1.00 ml

Vitamin B12 20.00 mkg

Distilled water 1000.00 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl2 × 4 H2O 1.5 g

ZnCl2 70.0 mg

MnCl2 × 4 H2O 100.0 mg

H3BO3 6.0 mg

CoCl2 × 6 H2O 190.0 mg

CuCl2 × 2 H2O 2.0 mg

NiCl2 × 6 H2O 24.0 mg

Na2MoO4 × 2 H2O 36.0 mg

Distilled water 990.0 ml

The trace element solution preparation: FeCl2 × 4 H2O is dissolved firstly in HCl, and then is mixed with water and other salts are dissolved in the sequence indicated.

Prepare and boil the medium in the absence of Na2S × 9 H2O, NaHCO3, vitamin B12, CaCl2 × 2 H2O, MgCl2x 6 H2O. Cool under a stream of nitrogen and add the remaining components, except vitamin B12 and Na2S2O3 × 5 H2O. Dispense into tubes or bottle fitted with rubber stoppers and autoclave. Add the vitamin B12 (filter sterilised) and Na2S2O3 × 5 H2O from sterile stock solutions.

Final pH 9.0 – 9.5

Autoclave at 121°C for 15 min.

 

Updated 25.06.2021