Medium data

Search for medium no. = [1004].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


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.

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