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 100ml of the medium (containing all additive but HEPES), pH should be around 7.6 at 30C. Otherwise, adjust pH of the HEPES solution with 10 N NaOH or conc. HCl. Finally fill up to 500 ml with distilled water.