Main sol. 815a

Equipment needed: Autoclave Hungate tubes Filter Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Na₂SO₄	3.0	g	3	21.121
CaCl₂ x 2 H₂O	6.0	g	6	40.812
NaCl	40.0	g	40	684.463
KCl	2.0	g	2	26.828
NH₄Cl	0.3	g	0.3	5.609
KH₂PO₄	0.2	g	0.2	1.47
MgCl₂ x 6 H₂O	8.0	g	8	39.35
SrCl2 x 6 H₂O	0.1	g	0.1	0.375
Trace element solution SL-11	1.0	ml	-	-
MOPS	3.0	g	3	14.336
Yeast extract	1.0	g	1	-
Sodium resazurin (0.1% w/v)	0.5	ml	5.0e-4	-
Na-pyruvate	2.5	g	2.5	22.718
Na₂CO₃	0.5	g	0.5	4.718
Na₂S x 9 H₂O	0.1	g	0.1	0.416
Distilled water	1000.0	ml	-	-
1 Dissolve ingredients except pyruvate, bicarbonate and sulfide, adjust pH to 6.0 with NaOH and sparge medium for 30 - 45 min with 100% N₂ gas to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add pyruvate (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Final medium pH should be 7.2.
2 Note: Prior to inoculation 10-20 mg/l sodium dithionite (added from a 5% w/v solution freshly prepared under N₂ and filter-sterilized) can be added to the medium to stimulate growth at the beginning.

815a	DESULFOCELLA HALOPHILA MEDIUM