Medium data

Search for medium no. = [1095].

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

1095 BICARBONATE BUFFERED MEDIUM

Na₂HPO₄·2H₂O	`0.53`	g
KH₂PO₄	`0.41`	g
NH₄Cl	`0.3`	g
NaCl	`0.3`	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)	`0.5`	ml
Yeast extract	`0.1`	g
Resazurin	`0.5`	mg
Distilled water	`930.0`	ml

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

10% MgCl₂·6H₂O solution	`1.0`	ml
10% CaCl₂·2H₂O solution	`1.0`	ml
Trace vitamins* (see Medium No. 197)	`10.0`	ml
8% NaHCO₃ solution*	`50.0`	ml
Substrate solution (see below)	`10.0`	ml

Aseptically and anaerobically distribute the medium into culture vessels (e.g., 50 ml of the medium in 120 ml serum bottle) under a N₂-CO₂ (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, add per liter the following solution (autoclaved and stored under a N₂ atmosphere):

5% Na₂S·9H₂O solution 10.0 ml

Substrate Solution:
For strain JCM 30818, 1 M glycerin solution;
For strain JCM 31247, 1 M trisodium citrate solution;
For strain JCM 31248, 1 M maltose solution.