Defined Minimal Medium for Vibrio fischeri
Final concentrations:
50 mM Tris
50 mM MgSO4
10 mM CaCl2
300 mM NaCl
10 mM KCl
0.333 mM K2PO4
18.5 mM NH4Cl *
32.6 mM Glycerol **
To make 1 liter:
500 ml Artificial Seawater Concentrate (see below)
50 ml of 1 M Tris-HCL (pH 7.2)
1 ml phosphate stock (see below)
5 ml ammonium stock (see below) *
3 g glycerol ** (for convenience, use 6 ml 50% glycerol w/v)
Build to 1 liter with tap water to supply trace metals
For liquid media, filter sterilize. The salts are likely to precipitate after autoclaving, so only do
so for solid media. Use 12 to 15 g agar per liter for plates.
* If the carbon source used contains nitorogen, the ammonium may be left out
** The glycerol can be replaced by a final concentration of 10 mM N-acetyl-glucosamine. If
using N-acetyl-glucosamine, do not add ammonium
2X Artificial Seawater concentrate (aka Defined Seawater)
Final concentrations:
100mM MgSO4
20 mM CaCl2
600 mM NaCl
20 mM KCl
To make (Note: Because some salts may form precipitates if mixed together directly, these
steps need to be followed in order and the salts dissolved separately)
Put 4 liters DI H2O into a 10-liter tank
Dissolve completely 198 g MgSO4-7H2O (or 96 g anhydrous MgSO4) in 1 liter of dH2O
Add resulting mixture to tank
Dissolve completely 23 g CaCl2-2H2O (or 17.5 g anhydrous CaCl2) in 1 liter of dH2O
Add to tank
Add 281 g NaCl and 12 g KCl directly to tank
Build total volume to 8 liters with dH2O
Stock nutrient solutions: Store sterile
Phosphate stock: 333 mM K2HPO4 (5.8% w/v)
Ammonium stock: 3.7 M NH4Cl (20% w/v)
Prepared by Sarah Studer July 2005
Modified from Ruby, Jan 2003