Medium data

Search for medium no. = [1339].

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

1339 MKD1 MEDIUM

MgCl₂·6H₂O	`9.47`	g
CaCl₂·2H₂O	`1.36`	g
NaCl	`20.75`	g
KCl	`0.6`	g
NH₄Cl	`0.54`	g
KH₂PO₄	`0.14`	g
NaHCO₃	`2.7`	g
Trace element solution (see Medium No. 1126)	`1.0`	ml
SrCl₂ solution (see below)	`10.0`	ml
NaF/LiCl solution (see below)	`10.0`	ml
Se/W solution (see Medium No. 852)	`2.0`	ml
Resazurin	`0.5`	mg
Distilled water	`1.0`	L

Mix components and autoclave. Cool down under a N₂-CO₂ gas stream. Aseptically and anaerobically add per liter the following solutions from sterile anaerobic stocks :

10% Casamino acid solution	`5.0`	ml
10% Powdered milk solution	`5.0`	ml
Vitamin solution (see below)	`1.0`	ml

Adjust pH to 7.5. Aseptically and anaerobically distribute the medium into culture vessels (e.g., 18 ml medium in 60 ml serum bottles), fill the head space with a gas mixture of N₂-CO₂ (4:1, v/v), and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution.

5% Na₂S·9H₂O solution	`3.3`	ml
5% L-Cysteine·HCl·H₂O solution	`3.3`	ml

SrCl₂ solution:

SrCl₂·6H₂O	`1.2`	g
Distilled water	`1.0`	L

NaF/LiCl solution:

NaF	`0.3`	g
LiCl	`0.1`	g
Distilled water	`1.0`	L

Trace vitamin solution:

Biotin	`4.9`	mg
Folic acid	`8.8`	mg
Pyridoxine-HCl	`4.1`	mg
Thiamine-HCl	`6.7`	mg
Riboflavin	`7.5`	mg
Nicotinic acid	`2.5`	mg
DL-Calcium pantothenate	`9.5`	mg
Vitamin B12	`0.1`	mg
p-Aminobenzoic acid	`2.7`	mg
Lipoic acid	`4.1`	mg
(R)-Pantoic acid sodium salt	`3.4`	mg
Distilled water	`1.0`	L

[Note]
It is modified from the trace vitamin solution of No. 284 by reducing vitamin B12 and by adding Pantoic acid. Casamino acid and powdered milk solutions could be replaced by 10% Yeast extract (BD-Difco) 5ml per liter.