Medium data

Search for medium no. = [1004].

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

1004 FRESHWATER MEDIUM FOR SOIL AOA

Basic FWM solution:

NaCl	`1.0`	g
KCl	`0.5`	g
MgCl₂·6H₂O	`0.4`	g
KH₂PO₄	`0.2`	g
CaCl₂·2H₂O	`0.1`	g
Distilled water	`1.0`	L

Autoclave Basic FWM solution and aseptically add the following solutions (autoclaved or *filter-sterilized) in the given order:

Modified trace element mixture (see below)	`1.0`	ml
Vitamin solution* (see below)	`1.0`	ml
7.5 mM EDTA·Na·Fe(III) solution* (pH 7.0)	`1.0`	ml
1 M NaHCO₃ solution*	`2.0`	ml
HEPES solution (see below)	`10.0`	ml
1 M NH₄Cl solution	`1.0`	ml
1 M Sodium pyruvate solution*	`1.0`	ml

Use glass or polystyrene containers for cultivation. Cultivate in the dark without shaking.
Modified trace element mixture:

HCl (25%)	`12.5`	ml
H₃BO₃	`30.0`	mg
MnCl₂·4H₂O	`100.0`	mg
CoCl₂·6H₂O	`190.0`	mg
NiCl₂·6H₂O	`24.0`	mg
CuCl₂·2H₂O	`2.0`	mg
ZnSO₄·7H₂O	`144.0`	mg
Na₂MoO₄·2H₂O	`36.0`	mg
Distilled water	`987.0`	ml

Vitamin solution:

Biotin	`20.0`	mg
Folic acid	`20.0`	mg
Pyridoxine·HCl	`100.0`	mg
Thiamine·HCl	`50.0`	mg
Riboflavin	`50.0`	mg
Nicotinic acid	`50.0`	mg
DL-Calcium pantothenate	`50.0`	mg
Vitamin B₁₂	`1.0`	mg
p-Aminobenzoic acid	`50.0`	mg
Choline chloride	`2.0`	g
Distilled water	`1.0`	L

Adjust pH to 7.0 with KOH.
HEPES solution:

NaOH	`12.0`	g
HEPES (free acid)	`119.2`	g

Dissolve NaOH in 300 ml distilled water. While stirring vigorously, slowly add HEPES until completely dissolved. Fill up to 450 ml with distilled water. If 1 ml of this solution is added to 100 ml of the medium (containing all additive but HEPES), pH should be around 7.6 at 30ºC. Otherwise, adjust pH of the HEPES solution with 10 N NaOH or conc. HCl. Finally fill up to 500 ml with distilled water.

[Note]
Adjust pH to 7.0 for JCM 31640.