Team:Groningen/protocols/biofilm media

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.

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