Team:Groningen/protocols/biofilm media
From 2013.igem.org
Make an overnight culture of the strains of interest in LB broth.
1 ml of the O/N culture is required for every 10 ml of Sgg medium (2x SG supplemented with 1% [wt/vol] glycerol).
This will be incubated at 27°C for 5 days.
1 ml of the O/N culture is required for every 10 ml of Sgg medium (2x SG supplemented with 1% [wt/vol] glycerol).
This will be incubated at 27°C for 5 days.
Media for macrocolony formation
2x SG medium (100 ml)
compound | amount | treatment |
---|---|---|
Nutrient broth (Difco) | 1.6 g | |
KCl | 0.2 g | |
MgSO4.7H2O | 0.05 g | |
agar | 1.5 g | |
add everything together and autoclave | ||
Ca(NO3)2.4H2O (1 mM) | 0.1 ml | |
MnCl2.4H2O (0.1 mM) | 0.1 ml | |
FeSO4 (1 µM) | 0.1 ml | |
0.1% glucose | 0.5 ml |
LB/Mn/glucose medium (100 ml)
compound | amount | treatment |
---|---|---|
LB/ 1.5% agar | 100 ml | autoclave |
MnCl2.4H2O (0.1 mM) | 0.1 ml | |
0.1% glucose | 0.5 ml |
MSgg medium (100 ml)
compound | amount | treatment |
---|---|---|
KH2PO4 | 0.026 g | |
K2HPO4 | 0.061 g | |
MOPS (100 mM) | 2.09 g | |
MgCl2.6H2O (2 mM) | 0.04 g | |
agar | 1.5 g | |
add everything together and autoclave | ||
CaCl2 (700 mM) | 0.1 ml | |
MnCl2 (50 µM) | 50 µl | |
FeCl3 (50 µM) | 0.1 ml | |
ZnCl2 (1 µM) | 0.1 ml | |
thiamine (2 µM) | 0.1 ml | |
0.5% glycerol | 0.57 ml | |
0.5% glutamate | 10 ml | |
tryptophan | 1 ml |