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DSMZ

DSMZ 816 . SPOROTOMACULUM MEDIUM

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Equipment needed: Autoclave Gassing station Hungate tubes
Final gas composition: 80 % N2 20 % CO2 read more
Final pH: 7.6
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ml
Compound Amount Unit
Solution A 920 ml
Solution B 1 ml
Solution C 1 ml
Solution D 50 ml
Solution E 10 ml
Solution F 5 ml
Solution G 5 ml
Solution H 10 ml
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B, C, F, G, and H are autoclaved separately under 100% N2 gas. Solution D is autoclaved under 80% N2 and 20% CO2 gas atmosphere and autoclaved. Solution E is prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be checked and adjusted to 7.6 with a sterile anoxic stock solution of Na2CO3 (5% w/v), if necessary.

Solution A #

920
Compound Amount Unit
KH2PO4 0.20 g
NH4Cl 0.30 g
KCl 0.50 g
NaCl 1.00 g
CaCl2 x 2 H2O 0.15 g
MgCl2 x 6 H2O 0.40 g
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 920.00 ml

Solution B #

1
Compound Amount Unit
Trace element solution SL-10 1 ml

Solution C #

1
Compound Amount Unit
Selenite-tungstate solution 1 ml

Solution D #

50
Compound Amount Unit
Na2CO3 2.5 g
Distilled water 50.0 ml

Solution E #

10
Compound Amount Unit
3-Hydroxybenzoic acid 2.8 g
Distilled water 10.0 ml
1 Neutralized with NaOH.

Solution F #

5
Compound Amount Unit
Yeast extract 0.5 g
Distilled water 5.0 ml

Solution G #

5
Compound Amount Unit
Trypticase
(BD BBL) 		0.5 g
Distilled water 5.0 ml

Solution H #

10
Compound Amount Unit
Na2S x 9 H2O 0.3 g
Distilled water 10.0 ml

Trace element solution SL-10 #

1000
Compound Amount Unit
HCl
(25%) 		10.0 ml
FeCl2 x 4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2 x 4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2 x 6 H2O 190.0 mg
CuCl2 x 2 H2O 2.0 mg
NiCl2 x 6 H2O 24.0 mg
Na2MoO4 x 2 H2O 36.0 mg
Distilled water 990.0 ml
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Selenite-tungstate solution #

1000
Compound Amount Unit
NaOH 0.5 g
Na2SeO3 x 5 H2O 3.0 mg
Na2WO4 x 2 H2O 4.0 mg
Distilled water 1000.0 ml