Medium data

Search for medium no. = [822].

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

822 FERMENTATIVE CELLULOLYTIC ANAEROBES MEDIUM

KCl	`0.33`	g
NH₄Cl	`0.25`	g
MgSO₄·7H₂O	`0.6`	g
NaCl	`0.4`	g
CaCl₂·2H₂O	`0.1`	g
Trace minerals (see Medium No. 151)	`10.0`	ml
Yeast extract	`0.2`	g
Resazurin	`0.25`	mg
1.0 M Phosphate buffer (see below)	`1.0`	ml
MOPS	`2.09`	g
Distilled water	`910.0`	ml

Mix components and bring to a boil for 5-10 sec, then, cool down under a N₂-CO₂ (80:20, v/v) gas stream. Dispense the medium into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add per liter the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized):

10% Cellobiose solution*	`40.0`	ml
Trace vitamins (see Medium No. 197)*	`10.0`	ml
5% NaHCO₃ solution*	`10.0`	ml
5% Na₂S·9H₂O solution	`10.0`	ml
5% L-Cysteine·HCl·H₂O	`10.0`	ml

1.0 M Phosphate buffer:
Mix 50 ml of 1.0 M KH₂PO₄ and 50 ml of 1.o M Na₂HPO₄ to prepare 100 ml of 1.0 M phosphate buffer.