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DSMZ

DSMZ 87 . CHLOROFLEXUS MEDIUM (modified)

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Equipment needed: Autoclave
Final pH: 8.2
Select a strain (optional):

ml
Compound Amount Unit
Yeast extract 1.00 g
Glycyl-glycine 1.00 g
Na2HPO4 x 2 H2O 0.10 g
MgSO4 x 7 H2O 0.10 g
KNO3 0.10 g
NaNO3 0.50 g
NaCl 0.10 g
CaCl2 x 2 H2O 0.05 g
Fe(III) citrate
(0.1 g in 100 ml H2O) 		5.00 ml
Trace element solution SL-6 1.00 ml
Distilled water 1050.00 ml
1 Adjust pH to 8.2.
2 Boil the medium under a stream of nitrogen gas for a few minutes and distribute 90 ml medium into 100 ml screw-capped bottles. Bubble each bottle with nitrogen gas and close immediately with a rubber septum and screw tight. Autoclave at 121°C for 15 min. After autoclaving inject 1.0 ml of neutralized sulfide solution (0.03% end concentration) to each bottle. This medium can be stored for several months. Incubate the culture at 50°C at a light intensity of 300 - 500 lux. For heavy cell suspension supplement periodically with sterile yeast extract solution (0.1% end concentration, for more details see Ref. 3365, 3366).

Neutralized sulfide solution 3% (w/v) #

100
Compound Amount Unit
Na2S x 9 H2O 3 g
Distilled water 100 ml
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle.

Trace element solution SL-6 #

1000
Compound Amount Unit
ZnSO4 x 7 H2O 0.10 g
MnCl2 x 4 H2O 0.03 g
H3BO3 0.30 g
CoCl2 x 6 H2O 0.20 g
CuCl2 x 2 H2O 0.01 g
NiCl2 x 6 H2O 0.02 g
Na2MoO4 x 2 H2O 0.03 g
Distilled water 1000.00 ml