Medium data

Search for medium no. = [770].

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

770 BETAINE+BICARBONATE BUFFERED MEDIUM

Solution A:

Phosphate solution (see below)	`15.0`	ml
Na₂SeO₃·5H₂O	`0.3`	mg
Na₂WO₄·2H₂O	`0.3`	mg
Yeast extract	`0.2`	g
Resazurin	`0.5`	mg
Distilled water	`900.0`	ml

Mix components thoroughly and autoclave under a N₂-CO₂ (80:20, v/v) gas mixture.
Solution B:

Trace metal solution (see below)	`1.0`	ml
Vitamin solution (see below)	`1.0`	ml
Mineral salt solution (see below)	`12.5`	ml
1.0 M Betaine solution	`10.0`	ml
Distilled water	`25.5`	ml

Mix components thoroughly and filter-sterilize. Stock the solution under a N₂-CO₂ (80:20, v/v) gas mixture.
Solution C:

NaHCO₃	`4.0`	g
Na₂S·9H₂O	`0.25`	g
L-Cysteine·HCl·H₂O	`0.5`	g
Distilled water	`50.0`	ml

Mix components thoroughly and filter-sterilize. Stock the solution under a N₂-CO₂ (80:20, v/v) gas mixture.
To complete the medium, combine Solutions A, B and C aseptically and anaerobically under a N₂-CO₂ (80:20, v/v) gas mixture. Check the final pH to be 6.9 - 7.2.
Phosphate solution:

KH₂PO₄	`0.41`	g
Na₂HPO₄	`0.43`	g
Distilled water	`100.0`	ml

Trace metal solution:

FeCl₂·4H₂O	`2.0`	g
H₃BO₃	`0.05`	g
ZnCl₂	`0.05`	g
CuCl₂·2H₂O	`0.03`	g
MnCl₂·4H₂O	`0.5`	g
Na₂MoO₄·2H₂O	`0.05`	g
AlCl₃	`0.05`	g
CoCl₂·6H₂O	`0.05`	g
NiCl₂·H₂O	`0.05`	g
EDTA	`0.5`	g
32% HCl	`1.0`	ml
Distilled water	`1.0`	L

Mineral salt solution:

NH₄Cl	`24.0`	g
NaCl	`24.0`	g
CaCl₂·2H₂O	`8.8`	g
MgCl₂·6H₂O	`8.0`	g
Distilled water	`1.0`	L

Vitamin solution:

Biotin	`0.02`	g
Folic acid	`0.02`	g
Pyridoxamine	`0.25`	g
Pyridoxine·HCl	`0.1`	g
Thiamine·HCl	`0.1`	g
Riboflavin	`0.05`	g
Nicotinic acid	`0.1`	g
Nicotinamide	`0.1`	g
DL-Calcium pantothenate	`0.05`	g
Vitamin B₁₂	`0.05`	g
p-Aminobenzoic acid	`0.05`	g
Lipoic acid	`0.05`	g
Distilled water	`1.0`	L