Medium data

Search for medium no. = [981].

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

981 ANAEROBIC BICARBONATE-BUFFERED MEDIUM WITH PHENOL

KH₂PO₄	`0.2`	g
NH₄Cl	`0.45`	g
NaCl	`1.7`	g
KCl	`0.5`	g
MgCl₂·6H₂O	`0.4`	g
CaCl₂·2H₂O	`0.16`	g
Na₂SO₄	`4.0`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Selenite tungstate solution (see Medium No. 431)	`1.0`	ml
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix components thoroughly and autoclave under a N₂-CO₂ (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized):

8.0% NaHCO₃ solution	`30.0`	ml
Vitamin solution (see Medium No. 403)	`1.0`	ml
Thiamine solution (see Medium No. 403)	`1.0`	ml
Vitamin B₁₂ solution (see Medium No. 403)	`1.0`	ml

Distribute the medium into sterile culture vessels (e.g., 50 ml per 120 ml bottle) under a N₂-CO₂ (4:1, v/v) gas mixture, and seal with sterile butyl rubber stoppers. Prior to inoculation, add per liter the following solutions (autoclaved or *filter-sterilized and stored under a N₂ atmosphere) in the order listed:

0.1 M Phenol solution* (freshly prepared)	`10.0`	ml
5% Na₂S·9H₂O solution	`10.0`	ml