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Reagents and Procedures For Pseudomonas Culturing: 1 M Mgso
Reagents and Procedures For Pseudomonas Culturing: 1 M Mgso
Reagents and Procedures For Pseudomonas Culturing: 1 M Mgso
1 M Tris·Cl
In a volumetric flask, dissolve 121 g/l Tris base; do not bring to volume yet. Adjust to desired pH
with concentrated HCl. Mix and bring to volume.
1 M CaCl2
Dissolve 11.10 g CaCl2 in 100 ml DI water.
Autoclave for 20 min.
Glycerol solution
65 ml glycerol
10 ml 1 M MgSO4
2.5 ml 1 M Tris·Cl, pH 8.0
22.5 ml DI water
Autoclave for 20 min.
Kings B Medium
Proteose Peptone 20 g 10 g 2g
Glycerol 10 g 5g 1g
K2HPO4 1.5 g 0.75 g 0.15 g
Agar (solid media only) 15 g 7.5 g 1.5 g
DI water 1 liter 500 ml 100 ml
Adjust to pH 7.2
Autoclave 20 min
Add sterile 1 M MgSO4
1 M MgSO4·7H2O 6.09 ml 3.05 ml 0.609 ml
Immediately before use:
For wild-type P. fluorescens CY091, add rifampin to a final concentration of 100 µg/ml
For P. fluorescens mutant J-1, add 100 µg/ml rifampin and 50 µg/ml kanamycin
Pipet 500 µl aliquots of the culture into labeled 1.2 ml freezer vials containing 500 µl glycerol
solution.