Medium data

Search for medium no. = [1389].

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

1389 IA91 MEDIUM

NH₄Cl	`0.54`	g
KH₂PO₄	`0.14`	g
MgCl₂·6H₂O	`3.0`	g
CaCl₂·2H₂O	`0.15`	g
NaCl	`20.0`	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
NaHCO₃	`2.5`	g
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix components, except NaHCO₃, and adjust pH 7.0. Bring to a boil for 5-10 seconds, then, cool down under a stream of N₂-CO₂ (80:20, v/v), add NaHCO₃, and autoclave under the same gas mixture. After cooling. aseptically and anaerobically, add the following solutions (autoclaved or *filter-sterilized, and stored under a N₂ atmosphere):

Trace vitamins* (see Medium No. 197)	`2.0`	ml
Culture supernatant (see below)	`400.0`	ml

Distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. Finally, add per liter the following solutions (autoclaved and stored under a N₂ atmosphere):

5% L-Cysteine·HCl·H₂O solution	`5.0`	ml
5% Na₂S·9H₂O solution	`5.0`	ml

Culture supernatant:
Centrifuge a grown culture of strain Acc8 (=JCM 39386) in JCM Medium No. 1388 (e.g., 10,000 xg, 10 min.), then, remove and autoclave the supernatant under a N₂-CO₂ (80:20, v/v) gas mixture.