Medium data

Search for medium no. = [695].

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

695 NAUTILIA ABYSSI MEDIUM

NaCl	`25.0`	g
NaNO₃	`1.0`	g
NH₄Cl	`0.33`	g
KCl	`0.33`	g
CaCl₂·2H₂O	`0.33`	g
MgCl₂·6H₂O	`0.33`	g
MES	`1.95`	g
Resazurin	`1.0`	mg
Trace minerals (see Medium No. 151)	`10.0`	ml
Sulfur (powder)	`12.0`	g
Distilled water	`1.0`	L

Mix components except sulfur, adjust pH to 6.0 and autoclave under a N₂ atmosphere. Distribute the medium into culture vessels (e.g., 20 ml in 100 ml serum bottles) containing appropriate amount of sulfur under a H₂-CO₂ (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized) into the medium (per liter):

Trace vitamins (see Medium No. 197)*	`10.0`	ml
2% KH₂PO₄ solution	`20.0`	ml
5% Na₂S·9H₂O solution	`10.0`	ml

Pressurize the inoculated culture vessel to 200 kPa H₂-CO₂ (4:1, v/v).