Medium data

Search for medium no. = [354].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

354 MODIFIED SULFATE REDUCING BACTERIA MEDIUM

NaCl	`20.0`	g
MgCl₂·6H₂O	`3.0`	g
Na₂SO₄	`4.0`	g
KH₂PO₄	`0.2`	g
KCl	`0.5`	g
NH₄Cl	`0.25`	g
PIPES	`3.46`	g
CaCl₂·2H₂O	`0.15`	g
Tryptone (BD-Difco)	`0.5`	g
Sodium acetate	`0.166`	g
Na₂WO₄·2H₂O	`2.0`	mg
Na₂SeO₄	`0.5`	mg
Trace vitamins (see Medium No. 197)	`10.0`	ml
Vitamin B₁₂	`0.05`	mg
Resazurin	`1.0`	mg
Na₂S·9H₂O	`0.5`	g
Distilled water	`1.0`	L

Mix ingredients, except Na₂S·9H₂O, and adjust pH to 6.7 with HCl. After autoclaving under a N₂ atmosphere, distribute the medium into sterile culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H₂-CO₂ (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Neutralize 5% Na₂S·9H2O solution, and autoclave under a N₂ atmosphere. Prior to inoculation, add the sterile Na₂S·9H₂O solution to the medium. Pressurize the inoculated vessels to 200 kPa H₂-CO₂ (4:1, v/v).